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AP-1 transcription factor

The AP-1 transcription factor is an inducible dimeric protein complex that regulates gene expression by binding to specific DNA sequences, such as the TPA-responsive element (TRE; 5′-TGAG/CTCA-3′) or cAMP-responsive element (CRE; 5′-TGACGTCA-3′), in response to extracellular stimuli. It consists of homo- or heterodimers formed by basic leucine zipper (bZIP) domain-containing proteins from the Jun (c-Jun, JunB, JunD), Fos (c-Fos, FosB, Fra-1, Fra-2), ATF (ATF2, ATF3, BATF), and Maf (c-Maf, MafA, MafB, MafG/F/K) families, enabling diverse functional specificities. First identified in the 1980s as a DNA-binding activity in mammalian cells and linked to proto-oncogenes like c-jun and c-fos discovered in retroviruses, AP-1 was formally characterized in the late 1980s through studies on phorbol ester-induced gene activation. AP-1's activity is primarily regulated by (MAPK) pathways, including ERK, JNK, and p38, which phosphorylate its subunits to enhance DNA binding, dimerization, and transactivation potential, often in concert with signaling. Additional control occurs through post-translational modifications, protein interactions, and of subunit expression, allowing context-specific responses to growth factors, cytokines, and signals. In physiological contexts, AP-1 governs essential cellular processes such as , , survival, , and , with particular importance in immune cell activation (e.g., T-cell cytokine production like IL-2 and IL-6) and epidermal . Dysregulation of AP-1 contributes to pathological conditions, exhibiting dual roles in cancer where subunits like c-Jun promote oncogenesis via and , while JunB and JunD often act as tumor suppressors. It also drives inflammatory responses and immune evasion mechanisms, such as upregulation of checkpoints like PD-1 and in tumors. These multifaceted properties position AP-1 as a key therapeutic target, with inhibitors explored for anticancer and applications.

Discovery and History

Initial Identification

The activator protein 1 (AP-1) transcription factor was first identified in the mid-1980s as a DNA-binding activity in nuclear extracts from phorbol ester (12-O-tetradecanoylphorbol-13-acetate, or TPA)-treated cells, responsive to this tumor-promoting agent. Initial studies revealed that this activity binds to specific enhancer elements in the promoter region of the human metallothionein IIA (hMT-IIA) gene, which exhibits high basal transcription levels and responsiveness to external stimuli. Concurrently, the same factor was found to interact with the enhancer of the simian virus 40 (SV40), a viral element implicated in oncogenic processes. These observations established AP-1 as a key mediator of TPA-inducible gene expression through biochemical assays, such as DNase I footprinting, which demonstrated protected regions in stimulated nuclear extracts from HeLa cells. The TPA-responsive element (TRE) was characterized as the consensus DNA sequence to which AP-1 binds, defined as 5’-TGAG/CTCA-3’, a palindromic motif conserved across multiple TPA-inducible promoters including those of hMT-IIA, SV40, collagenase, and stromelysin. Synthetic oligonucleotides containing this TRE conferred TPA inducibility to heterologous promoters in transfection assays, confirming its functional role as an enhancer. Purification of AP-1 to near homogeneity from HeLa cell nuclear extracts via sequence-specific DNA affinity chromatography yielded a 47 kDa polypeptide that specifically activated transcription from wild-type hMT-IIA templates but not from mutants lacking the TRE. TPA treatment enhanced AP-1 binding activity 3- to 4-fold in nuclear extracts through a posttranslational mechanism, without altering the abundance of the factor itself. These early discoveries linked AP-1 activity to the regulation of genes involved in cellular responses to tumor promoters, providing an initial connection to oncogene activation via SV40 enhancer interactions. Subsequent work identified the protein components of AP-1, such as c-Fos and c-Jun, but the foundational biochemical characterization established its role as a stimulus-responsive .

Key Milestones and Oncogenic Insights

In late 1987 and early 1988, the of the c-jun proto-oncogene marked a pivotal advancement in understanding AP-1, as it identified the cellular homolog of v-Jun, the oncoprotein encoded by avian sarcoma virus 17 (ASV-17). Angel et al. isolated the human c-jun cDNA and demonstrated that its product functions as a sequence-specific transcriptional activator with properties akin to the previously characterized AP-1 factor, establishing a direct link between viral oncogenesis and cellular transcription regulation. In December 1987, Bohmann et al. cloned the human c-jun gene and showed that it encodes a structurally and functionally related to AP-1, confirming v-Jun's role as the transforming agent in ASV-17-transformed cells and highlighting AP-1's involvement in oncogenic signaling. That same year, the identification of the c-fos proto-oncogene further illuminated AP-1's oncogenic connections. Curran and Franza characterized c-Fos as the cellular counterpart to v-Fos, the oncoprotein from FBJ murine osteosarcoma virus, and proposed that Fos and Jun proteins form a heterodimeric complex responsible for AP-1 activity, thereby integrating immediate-early gene expression with tumor-inducing mechanisms. This revelation underscored how retroviral oncogenes like v-fos disrupt normal cellular control by hijacking AP-1-mediated transcription. By the late 1980s and early 1990s, research established that AP-1 operates as a dimeric rather than a monomeric entity, expanding its regulatory scope. Halazonetis et al. demonstrated that c-Jun can homodimerize or form heterodimers with c-Fos, yielding complexes with varying DNA-binding affinities to the TPA-responsive element (TRE), a initially identified in promoter analyses of phorbol ester-inducible genes. This dimeric nature, revealed through biochemical and genetic studies, provided key insights into AP-1's versatility in transducing extracellular signals to changes implicated in oncogenesis. The 1990s brought broader recognition of AP-1's role in via immediate-early genes, with foundational work on cellular responses to stimuli.

Molecular Structure

Protein Families and Components

The AP-1 transcription factor is primarily composed of dimeric complexes formed by proteins from the and Fos families, which belong to the basic (bZIP) class of transcription factors. The family includes three main members: c-Jun, JunB, and JunD. These proteins share a high degree of structural similarity, particularly in their bZIP domains, which enable DNA binding and protein-protein interactions. All family members are capable of forming homodimers among themselves or heterodimers with proteins from other families, allowing for diverse . c-Jun, the prototypical member, was identified as a key component of AP-1 and exhibits potent potential, while JunB and JunD often act as modulators with weaker or context-dependent activity. The Fos family consists of four proteins: c-Fos, , Fra-1 (fos-related antigen 1), and Fra-2. Unlike the Jun proteins, Fos family members lack the ability to homodimerize and instead form stable heterodimers exclusively with proteins to constitute functional AP-1 complexes. c-Fos, the founding member, is particularly notable for its role in rapid transcriptional responses, while , Fra-1, and Fra-2 contribute to sustained or specialized AP-1 activity, with Fra-1 and Fra-2 often displaying greater stability in chronic conditions. These heterodimers bind to TPA-responsive elements (TREs) in DNA, facilitating control. In addition to the core Jun and Fos families, AP-1 can incorporate proteins from other bZIP families to form specialized heterodimers, expanding its functional repertoire. The ATF family, particularly ATF2 (also known as CREB-p1), can heterodimerize with proteins to bind CRE/ATF sites, influencing responses to and signals. Similarly, Maf family members such as c-Maf, MafB, MafA, MafG, and MafK form heterodimers with Jun or Fos to regulate tissue-specific genes, often with repressive or activating effects depending on the context. The JDP family, including JDP1 and JDP2, acts predominantly as inhibitors by forming inactive heterodimers with Jun or c-Fos, thereby fine-tuning AP-1 activity. Members of the Jun and Fos families are classified as immediate-early genes, characterized by their rapid transcriptional induction in response to extracellular stimuli without requiring protein synthesis. Their mRNAs exhibit short half-lives, typically on the order of 10-20 minutes for c-Fos and around 11 minutes for c-Jun, ensuring transient expression that allows quick adaptation to changing cellular environments. Protein stability varies, with c-Fos having a half-life of approximately 2 hours and c-Jun around 90 minutes, contributing to the dynamic nature of AP-1 complexes. These expression patterns underscore the role of AP-1 components in orchestrating immediate cellular responses. Dimerization occurs via the motif, enabling combinatorial diversity in AP-1 function.

Dimerization and DNA Binding

The AP-1 transcription factor functions through dimeric complexes formed by members of the Jun, Fos, and ATF protein families, which share a conserved basic leucine zipper (bZIP) domain responsible for both dimerization and DNA binding. The bZIP domain consists of two structural motifs: a basic region rich in positively charged amino acids that facilitates sequence-specific contact with DNA, and a leucine zipper region characterized by a heptad repeat of leucine residues that mediates coiled-coil dimerization between two protein monomers. This architecture allows AP-1 proteins to assemble as homodimers or heterodimers, enabling versatile regulation of target genes.90147-X) Dimerization preferences among AP-1 components determine their DNA-binding specificity and efficiency. Jun family proteins, such as c-Jun, can form homodimers that bind to the palindromic 12-O-tetradecanoylphorbol-13-acetate-responsive element (TRE; consensus sequence TGAGTCA), while heterodimerization with Fos family proteins, such as c-Fos, preferentially targets the same TRE motif but with enhanced stability. In contrast, heterodimers between Jun and ATF/CREB family proteins, such as ATF-2, exhibit a preference for cyclic AMP response element (CRE)-like sites (consensus ATGACGTC), allowing AP-1 to interact with a broader repertoire of regulatory elements. These preferences arise from compatibility in the leucine zipper interfaces, where Fos lacks the ability to homodimerize effectively due to its zipper sequence, thus relying on Jun for complex formation.90147-X) Variations in DNA-binding are influenced by dimer and the of the target site. Jun/Fos heterodimers typically exhibit higher for the TRE than Jun/Jun homodimers, with binding efficiencies reported up to 25-fold greater, attributed to optimal spacing of the half-sites in the 7-base pair TRE core (TGAGTCA) that aligns the regions for cooperative interaction.90147-X) This enhanced in heterodimers stems from more stable dimerization and precise positioning on the DNA, whereas homodimers may require adjustments in half-site spacing for effective binding. For /ATF heterodimers, for CRE-like sites is similarly elevated compared to individual homodimers, reflecting adaptations in the region that accommodate the slightly divergent half-site orientation in CRE sequences. The molecular model of AP-1 DNA binding involves the leucine zipper forming a Y-shaped coiled-coil structure that positions the two basic regions to straddle the DNA helix. Upon dimerization, the basic regions extend as alpha-helices that insert into the major groove of the DNA, making direct contacts with the phosphate backbone and nucleotide bases of the TRE or CRE half-sites to achieve sequence specificity.90147-X) The zipper domain stabilizes this configuration by hydrophobic interactions between leucine residues, preventing dissociation and ensuring the fork-like grip on the DNA double helix. This binding mode induces a localized conformational change in the DNA, facilitating transcriptional activation while maintaining the integrity of the dimer interface.

Evolutionary Conservation and Crystal Structures

The basic leucine zipper (bZIP) domain central to AP-1 function exhibits high evolutionary conservation, spanning from unicellular eukaryotes to mammals, with the Gcn4 serving as a functional homolog that binds similar consensus sequences as AP-1 sites. This preservation underscores the domain's critical role in dimerization and DNA recognition, with sequence identity in key hydrophobic and basic residues maintained across distant taxa. Networks of bZIP interactions have diversified over a billion years while retaining core structural motifs from to humans. Orthologs of the core AP-1 components and Fos are evident in key models. In , the Jun ortholog DJra (also called Jra) and Fos ortholog (Kay) form heterodimers that mediate JNK signaling and developmental processes analogous to mammalian AP-1. Similarly, in , jun-1 and fos-1 encode bZIP proteins that dimerize to regulate reproductive and stress responses, mirroring AP-1's DNA-binding specificity and functional partnerships. These invertebrate counterparts highlight the ancient origins of AP-1-like regulation in metazoan evolution. Non-canonical AP-1 family members, such as those in the JDP subfamily, display more variable conservation, with homologs identified in some invertebrates like moths but absent or diverged in others, suggesting later evolutionary expansion in vertebrates. High-resolution structural studies have elucidated the atomic details of AP-1 dimerization and DNA interaction. The seminal 3.0 Å crystal structure of the c-Fos/c-Jun bZIP heterodimer bound to DNA (PDB: 1FOS) revealed parallel α-helices forming a leucine zipper that clamps the DNA major groove, with basic regions inserting into adjacent half-sites of the AP-1 consensus sequence. This 1995 structure demonstrated the asymmetric interface stabilizing the heterodimer over homodimers. For the c-Jun homodimer, an NMR solution structure of the leucine zipper domain (PDB: 1JUN) illustrated the symmetrical coiled-coil arrangement, while a later 2.2 Å crystal structure of the full bZIP domain bound to CRE DNA (PDB: 1JNM) confirmed similar helical continuity and groove-spanning contacts. Post-2000 structural analyses expanded insights into AP-1 variants and dynamics. A 3.0 Å of the ATF2/c-Jun heterodimer bound to the interferon-β enhancer (PDB: 1T2K) highlighted interdomain contacts beyond the bZIP core, including ATF2's motif contributing to with IRF-3. Complementary NMR studies of bZIP dimers, such as the c-Jun , revealed intrinsic flexibility in linker regions adjacent to the zipper, allowing conformational adaptability in non-bZIP activation domains during . These findings emphasize the structural enabling AP-1's diverse partnerships and responses to cellular signals.

Activation and Regulation

Upstream Signaling Pathways

The activation of AP-1 transcription factors is primarily triggered by extracellular signals that converge on intracellular kinase cascades, particularly the (MAPK) pathways, leading to the and of AP-1 components such as c-Jun, c-Fos, and ATF2. These signals include growth factors, cytokines, and stress stimuli that initiate rapid cellular responses. The c-Jun N-terminal kinase (JNK) pathway plays a central role in AP-1 activation by phosphorylating c-Jun at serine residues 63 and 73, enhancing its transcriptional activity in response to stress and inflammatory signals. JNK is activated downstream of various receptors, such as those for tumor necrosis factor-α (TNF-α), where prolonged JNK activity leads to sustained AP-1 stimulation. Similarly, the extracellular signal-regulated kinase (ERK) pathway, activated by mitogenic stimuli, induces c-Fos expression through phosphorylation of the ternary complex factor Elk-1, which binds to the serum response element in the c-fos promoter. The p38 MAPK pathway contributes by phosphorylating ATF2, a partner in AP-1 heterodimers, particularly in response to environmental stresses. Beyond MAPKs, other pathways influence AP-1, including (PKC) activation by phorbol esters like 12-O-tetradecanoylphorbol-13-acetate (TPA), which directly stimulates AP-1 binding to TPA-responsive elements. Crosstalk with occurs in inflammatory contexts, where can modulate JNK activity to enhance AP-1 function, amplifying proinflammatory . Growth factor receptors, such as the (EGFR), activate the Ras-ERK cascade to promote AP-1-dependent proliferation signals. Cytokine receptors, exemplified by TNF-α receptors, engage JNK to drive AP-1 activation. AP-1 activation exhibits temporal dynamics characteristic of immediate-early responses, with induction occurring within minutes of stimulation, enabling rapid adaptation to environmental cues before protein synthesis is required. This swift onset distinguishes AP-1 from delayed responses, facilitating its role in coordinating early transcriptional events.

Post-Translational Modifications

Post-translational modifications (PTMs) play a crucial role in regulating the stability, localization, and transcriptional activity of AP-1 transcription factors, enabling rapid responses to extracellular signals. These covalent changes, including , ubiquitination, , sumoylation, and modifications, modulate AP-1 dimer function without requiring protein synthesis. Phosphorylation is one of the most prominent PTMs affecting AP-1, particularly targeting the N-terminal of c-Jun. c-Jun N-terminal (JNK) phosphorylates c-Jun at serine residues 63 and 73, which enhances its potential and promotes of coactivators to target promoters. This modification is essential for AP-1 activation in stress responses, as unphosphorylated c-Jun exhibits reduced transcriptional efficacy. In contrast, glycogen synthase 3β (GSK3β) phosphorylates c-Jun at threonine 239 within its , inhibiting its activity and priming it for degradation to prevent prolonged signaling. Ubiquitination controls AP-1 via the , ensuring transient activity. The E3 ubiquitin ligase FBW7 recognizes GSK3β-phosphorylated c-Jun and promotes its polyubiquitination, leading to rapid proteasomal and termination of AP-1 signaling. Similarly, c-Fos undergoes ubiquitin-mediated destabilization, with its C-terminal serving as a that facilitates proteasomal breakdown, thereby limiting the duration of inducible AP-1 complexes. Certain non-degradative ubiquitination events, such as K63-linked chains, can protect AP-1 components from proteasomal under , stabilizing dimers for sustained activity. Acetylation and sumoylation provide additional layers of by altering AP-1 interactions with and other factors. The acetyltransferases CBP and p300 acetylate c-Jun at 271 in its , enhancing its transcriptional potency by facilitating coactivator binding and acetylation at target loci. Conversely, JunB is sumoylated at 237, repressing its function and shifting AP-1 toward inhibitory dimers in anti-proliferative contexts. Redox modifications, driven by (ROS), directly impact AP-1 DNA binding. Oxidation of conserved residues in the basic DNA-binding region of and Fos proteins forms intramolecular disulfide bonds, inhibiting TRE recognition and suppressing AP-1-dependent transcription during . This reversible allows AP-1 to integrate signals, with reduction by restoring binding activity.

Interactions with Co-Regulators

AP-1 s interact with various co-activators to enhance accessibility and transcriptional initiation at target promoters. The co-activators (CBP) and its paralog p300 bind to the of c-Jun, a core AP-1 component, facilitating recruitment to AP-1 binding sites and subsequent . This interaction promotes acetylation of H3 and H4 at promoter regions, loosening structure and enabling efficient transcription factor assembly and (Pol II) engagement. Seminal studies demonstrate that CBP/p300's (HAT) activity is essential for AP-1-mediated activation of genes involved in and stress responses, with disruption of this binding impairing transcriptional output. The complex further amplifies AP-1's transcriptional potency by bridging AP-1 dimers to the Pol II pre-initiation complex. AP-1 recruits subunits, such as MED1, to enhancers and promoters, stabilizing Pol II recruitment and of its C-terminal domain to transition from pausing to productive elongation. This co-activator function is particularly evident in signal-induced , where AP-1-guided assembly coordinates with general transcription factors to boost Pol II processivity at immediate-early genes like c-fos. In contrast, co-repressors such as (NCoR1) and silencing mediator for retinoid and thyroid hormone receptors (SMRT) suppress AP-1 activity through recruitment of histone deacetylases (HDACs). NCoR1/SMRT complexes bind unphosphorylated c-Jun at AP-1 sites, docking HDAC3 to deacetylate s and condense , thereby silencing inflammatory gene networks in macrophages and other immune cells. This HDAC-mediated repression integrates anti-inflammatory signals, preventing excessive AP-1-driven proinflammatory responses during . The Groucho/TLE family of co-repressors, including TLE1, modulates AP-1 output by interacting with JunB to attenuate transcription at select promoters. TLE proteins are recruited via short motifs in JunB, recruiting HDACs and promoting compaction to repress genes involved in and control. This repression is context-dependent, often counterbalancing activating AP-1 dimers in developmental and oncogenic settings. Chromatin remodeling complexes like also cooperate with AP-1 to overcome barriers. As a pioneer factor, AP-1 binds closed and recruits the BAF variant of via interactions with subunits like ARID1A and SMARCC1, driving ATP-dependent eviction and enhanced DNA accessibility at enhancers. This facilitates AP-1's access to embedded binding sites, reshaping 3D landscapes for sustained transcriptional activation during epithelial-mesenchymal transitions and stress . Over 90% of AP-1 peaks colocalize with occupancy, underscoring their interdependent roles in dynamic gene regulation. AP-1 engages in with other regulators to fine-tune responses to cellular cues. In , AP-1 cooperates with Nrf2 at composite promoter elements, such as in the sulfiredoxin gene (Srx), where both factors synergistically drive expression of enzymes to mitigate damage. This interaction enhances cytoprotection without competing for binding sites, integrating AP-1's proliferative signals with Nrf2's detoxifying program. Similarly, AP-1 intersects with β-catenin in Wnt signaling, where c-Jun physically associates with β-catenin/TCF complexes at promoters, amplifying transcription of shared targets like c-Myc and to promote and tumorigenesis. This occurs via direct protein-protein contacts, enabling Wnt to potentiate AP-1 activity in intestinal and cancer contexts, though it can also lead to mutual antagonism depending on signaling context. Dimer composition influences co-regulator preference, with Fos-Jun heterodimers favoring CBP/p300 over repressors like NCoR.

Biological Functions

Cell Proliferation and Senescence

The AP-1 transcription factor complex, particularly dimers composed of c-Jun and Fos family members, plays a pivotal role in promoting by directly inducing the expression of key regulators such as , cyclin A, and cyclin-dependent kinases (CDKs). These targets facilitate the G1-to-S phase transition, enabling progression through the in response to mitogenic signals. For instance, c-Jun/Fos heterodimers bind to AP-1 sites in the promoter, driving its transcription and subsequent activation of CDK4/6 complexes that phosphorylate the , thereby releasing transcription factors to promote S-phase entry. In contrast, certain AP-1 subunits exert inhibitory effects on ; notably, JunB acts as a negative regulator by repressing c-Jun-dependent targets, including through direct transcriptional activation of the inhibitor ^INK4a, which enforces G1 arrest. This antagonistic function of JunB highlights the context-dependent nature of AP-1 activity in balancing proliferative signals. AP-1 also contributes to , a state of permanent cell cycle arrest, particularly in oncogene-induced (). Fra-1/JunD dimers are key mediators, upregulating senescence effectors like p16^INK4a and p21^CIP1 to impose G1 arrest and remodel the landscape at senescence-associated enhancers. These dimers pioneer enhancer accessibility, facilitating the expression of () factors such as IL1A and IL1B, which reinforce the senescent state. Studies in RAS-driven models demonstrate that perturbing AP-1 activity, including Fra-1 and JunD, can reverse this transcriptional program, underscoring its reversible nature. Experimental evidence from models supports these roles; for example, conditional inactivation of AP-1 factors in the leads to basal hyperproliferation and disrupted control, manifesting as thickened layers and delayed .

Differentiation and Development

The AP-1 transcription factor complex, particularly involving c-Jun and c-Fos, plays a pivotal role in keratinocyte terminal by activating the promoters of differentiation-specific markers such as keratins and K10. In primary human keratinocytes, overexpression of c-Jun and c-Fos transactivates the K1 promoter through AP-1 binding sites, while c-Jun alone enhances K10 promoter activity, thereby promoting the expression of these suprabasal keratins essential for epidermal barrier formation. This activation occurs in response to differentiation cues like elevated calcium levels, where AP-1 integrates signals from the JNK pathway to drive commitment to the differentiated state. Conversely, disruption of AP-1 function, such as through dominant-negative c-Jun (TAM67) expression in suprabasal epidermis, impairs the upregulation of late differentiation genes, underscoring AP-1's necessity for completing the keratinization program. In bone remodeling, AP-1 dimers containing Fra-2 and JunD maintain the balance between osteoblast and osteoclast activities by positively regulating osteoblast differentiation and extracellular matrix production. Conditional knockout of Fra-2 in osteoblast lineage cells of mice results in severe osteopenia due to defective osteoblast maturation, reduced collagen type I synthesis, and impaired mineralization, highlighting Fra-2/AP-1 as a critical driver of bone formation. Similarly, JunD promotes osteoblast differentiation by antagonizing inhibitory factors like menin, with JunD-deficient mice exhibiting enhanced osteoblast activity and increased bone mass, demonstrating its role in fine-tuning osteoblast-osteoclast coupling. These findings from genetically modified mice reveal AP-1's context-dependent regulation of bone cell fate, where specific dimers ensure coordinated remodeling without overlapping proliferative effects. During mouse embryonic development, AP-1 components such as c-Jun and JunB are essential for and limb bud formation, often integrating signaling to pattern tissues. In c-Jun knockout embryos, defects in neural crest-derived structures arise from impaired , as c-Jun mediates JNK-dependent and motility in response to gradients along the . while in limb buds, c-Jun-containing AP-1 dimers regulate interdigital and patterning via -responsive elements, ensuring proper anterior-posterior axis formation. These roles position AP-1 as a downstream effector of signaling in orchestrating embryonic fate decisions and morphogenetic movements. AP-1 family members, including c-Fos, are crucial for placental invasion and uterine implantation in mice, facilitating the maternal-fetal interface. Although c-Fos knockout mice are viable, expression of c-Fos in cells supports invasive behavior during early , with its induction correlating with implantation window preparation in the . More prominently, related AP-1 proteins like Fra-1 (Fosl1) are indispensable, as Fosl1-null embryos fail to undergo proper invasion into uterine spiral arteries, resulting in defective vascular remodeling and mid-gestational due to impaired implantation and exchange. Dimer-specific activities, such as c-Fos/c-Jun complexes, further modulate motility by regulating expression essential for decidual penetration.

Apoptosis and Stress Response

The AP-1 transcription factor plays a dual role in , promoting in certain contexts while inhibiting it in others, depending on the cellular environment and dimer composition. In pro-apoptotic scenarios, c-Jun, a core component of AP-1, drives the expression of (FasL), which activates the extrinsic apoptosis pathway following (UV) irradiation. This induction occurs through JNK-mediated of c-Jun, enabling AP-1 binding to the FasL promoter and triggering caspase-dependent cell death in and fibroblasts. Similarly, c-Jun upregulates the BH3-only protein Noxa, a target that sensitizes mitochondria to pro-apoptotic signals, thereby enhancing UV-induced in cells. These mechanisms highlight c-Jun's role in eliminating damaged cells to prevent . The JNK-AP-1 axis is particularly critical in neuronal , where sustained JNK activation phosphorylates c-Jun, leading to transcriptional induction of pro-death genes. In sympathetic neurons deprived of , c-Jun deficiency blocks , demonstrating its essential function downstream of JNK in cytochrome c release and activation. This pathway operates independently of in some neuronal models, underscoring AP-1's direct contribution to developmental and stress-induced neuronal elimination. Conversely, AP-1 can exert anti-apoptotic effects through specific dimers, such as Fra-1-containing complexes that upregulate anti-apoptotic members. In transformed cancer cells, Fra-1 binds to the promoter, elevating its expression and inhibiting mitochondrial outer membrane permeabilization, thereby promoting cell survival under genotoxic stress. This protective function is evident in and models, where Fra-1 overexpression correlates with resistance to chemotherapy-induced death. In the stress response, AP-1 dimers involving ATF2 and proteins coordinate adaptation to DNA damage by synergizing with p53. The /ATF2 heterodimer binds to AP-1 sites in the promoters of genes like GADD153, enhancing p53-dependent transcription and facilitating arrest or repair rather than immediate . This synergy is activated by /ATR kinases following genotoxic insults, allowing cells to prioritize survival. Additionally, c-Fos contributes to heat shock responses by inducing heat shock protein 70 (), which chaperones misfolded proteins and prevents aggregation during . The context-dependence of AP-1 in and stress arises from dimer switching, where JunD-containing complexes provide protection against . Unlike pro-apoptotic c-Jun homodimers, JunD/Fos heterodimers upregulate genes such as MnSOD, reducing (ROS) levels and preventing endothelial and damage in models of vascular injury. This protective role is further supported by post-translational modifications, such as , that modulate ROS-sensitive dimerization.00804-9)

Physiological Roles

Tissue-Specific Regulation

In the liver, AP-1 activity, particularly through dimers involving JunB and Fra-1, contributes to regeneration following injury such as partial hepatectomy or toxin exposure. These dimers are induced as part of the immediate-early gene response, supporting and survival during the regenerative process by modulating genes involved in and anti-apoptotic pathways. For instance, Fra-1 overexpression enhances protection against acetaminophen-induced injury by upregulating glutathione S-transferase P1 (GSTP1) in hepatocytes, thereby facilitating recovery. In the skin, c-Jun plays a pivotal role in AP-1-mediated responses to injury and environmental stress, driving migration and proliferation essential for and UV-induced repair. During , c-Jun promotes re-epithelialization by regulating genes that facilitate epithelial cell movement across the wound bed, and its conditional in results in delayed closure and impaired re-epithelialization due to defective . Similarly, UV rapidly activates c-Jun, leading to AP-1 dimer formation that initiates protective responses like remodeling and resolution in epidermal cells. In the , AP-1 complexes formed by ATF3 and Jun family members, such as , are critical for neuronal survival in response to ischemic injury. Following cerebral ischemia, ATF3 and phosphorylated are upregulated in neurons, where they cooperatively induce heat shock protein 27 (Hsp27) expression, promoting anti-apoptotic mechanisms and regeneration to mitigate . ATF3 deficiency exacerbates neuronal post-ischemia, underscoring its protective function in this context via interaction with c-Jun pathways. In the heart, c-Fos activation within AP-1 dimers is integral to pathological hypertrophy triggered by angiotensin II signaling. Angiotensin II rapidly induces c-fos expression in cardiac myocytes through G-protein-coupled receptor pathways, leading to AP-1 binding that drives hypertrophic gene programs and cell enlargement. This response contributes to adaptive remodeling under hemodynamic stress, with c-Fos serving as a key immediate-early mediator in the transition to hypertrophy. Tissue-specific regulation of AP-1 often involves preferential dimer compositions, such as JunB/Fra-1 in the liver or c-Jun/ATF3 in the , reflecting local isoform expression patterns that fine-tune responses to .

Role in Immune Response and Inflammation

AP-1 transcription factors, particularly the c-Jun/c-Fos heterodimers, play a central role in production during T-cell activation. Upon (TCR) and co-stimulation, these dimers bind to TPA-responsive elements (TREs) in the promoters of interleukin-2 (IL-2) and tumor factor-alpha (TNF-α), driving their transcription. Specifically, c-Jun by JNK enhances AP-1 binding to composite NFAT/AP-1 sites in the IL-2 promoter, essential for T-cell proliferation and effector function. Similarly, AP-1 activation via ERK and JNK pathways induces TNF-α expression in T-cells, contributing to inflammatory signaling. This regulation underscores AP-1's integration with upstream signaling, such as the JNK pathway in (TLR) responses, to orchestrate adaptive immune activation. In macrophages, AP-1 components like Fra-1 promote pro-inflammatory activation and polarization. Fra-1 binds to the IL-6 promoter in response to stimuli such as (LPS), enhancing IL-6 production that acts autocrine to drive macrophage skewing toward inflammatory phenotypes. Although primarily linked to alternative activation in some contexts, Fra-1 contributes to the expression of inducible (iNOS) and other M1-associated genes by facilitating AP-1-mediated transcription of pro-inflammatory mediators during innate immune responses. This mechanism amplifies production and clearance but can perpetuate if dysregulated. AP-1 hyperactivity is implicated in autoimmune conditions, notably (RA), where elevated c-Jun activity in synovial fibroblasts sustains . Nuclear extracts from RA synovial tissues exhibit significantly higher AP-1 DNA-binding compared to controls, correlating with increased expression of matrix metalloproteinases and that drive joint destruction. The AP-1 family member JunD promotes inflammatory activation in . In JunD-deficient macrophages, pro-inflammatory —including TNF-α and IL-6—is reduced, leading to dampened activation and cytokine secretion. This function of JunD supports macrophage responses in immune .

Involvement in Cancer and Other Diseases

The AP-1 transcription factor exhibits paradoxical roles in cancer, acting as both an and tumor suppressor depending on the cellular context and family member involved. In , particularly non-small cell lung cancer (NSCLC), c-Jun promotes tumorigenesis through activation of downstream pathways that enhance cell survival and resistance to therapies such as . Overexpression of c-Jun has been linked to aggressive phenotypes in NSCLC, where it drives metabolic reprogramming and glutaminase expression to support tumor growth. Similarly, in , Fra-1 (encoded by FOSL1) facilitates invasion by inducing matrix metalloproteinase 9 (MMP9) expression, enabling degradation and ; elevated Fra-1 levels correlate with poor prognosis in subtypes. Conversely, certain AP-1 components exert tumor-suppressive effects. Loss or downregulation of JunB in B-lymphoid cells promotes and , contributing to development, as evidenced by surveys of human samples showing reduced JunB expression in aggressive cases. In anaplastic large cell (ALCL), deletion of JunB alongside c-Jun exacerbates tumor progression by altering signaling pathways like PDGFRβ. These findings highlight JunB's role in restraining oncogenic signaling, including induction during . Beyond cancer, AP-1 dysregulation contributes to non-oncologic diseases. In , activation of c-Jun/AP-1 in drives hyperproliferation and production, exacerbating skin lesions; Fra-1 overexpression further shifts toward an epithelial-mesenchymal transition-like state. In neurodegeneration, such as (ALS), upregulated c-Jun in motor neurons promotes axonal degeneration and autophagic responses, with phosphorylated c-Jun observed in affected tissues. Therapeutic strategies targeting AP-1 have advanced, particularly for disease-associated dysregulation. The selective AP-1 inhibitor T-5224, which blocks c-Fos/c-Jun dimers, has shown preclinical efficacy in reducing by suppressing proinflammatory gene expression and is under investigation for inflammatory conditions like . Post-2020 developments include proteolysis-targeting chimeras (PROTACs) designed to degrade AP-1 family members; for instance, T-5224-based PROTACs selectively degrade Fra-1 (FOSL1) in head and neck , reducing cancer stemness without broad toxicity. These approaches underscore AP-1's potential as a druggable target in cancer and .

Target Genes and Regulome

Consensus Binding Sites and Motifs

The AP-1 transcription factor, composed of bZIP domain-containing proteins such as and Fos family members, primarily recognizes palindromic DNA sequences known as TPA-responsive elements (TREs). The TRE consensus sequence is 5'-TGAGTCA-3', a 7-base pair identified in the promoters of phorbol ester-inducible genes, including the human (MMP-1) gene. This sequence allows for symmetric binding by AP-1 dimers, with each contacting a half-site centered on the invariant TGAC core. Variants of the exist, influenced by the specific dimer composition. For instance, /ATF heterodimers preferentially bind to CRE-like sequences, such as 5'-ATGACGTCAT-3', which features an additional central pair compared to the TRE, enabling recognition by ATF/CREB family proteins in complex with . In contrast, Fos/ heterodimers exhibit higher affinity for the strict TRE , while homodimers show broader tolerance but reduced efficiency on TRE compared to CRE sites. Flanking sequences adjacent to the core modulate binding affinity; for example, -rich contexts enhance specificity and stability for Fos/ dimers by influencing DNA shape and bendability. Non-consensus AP-1 sites, which deviate from the ideal TRE, can still support dimer binding but often with altered specificity. In the promoter of the TIMP-1 gene, a variant sequence 5'-TGAGTAA-3' serves as a functional AP-1 site, where the terminal A substitution reduces affinity for classical Fos/Jun but accommodates other dimers under specific signaling conditions. In vitro selection methods, such as SELEX, have confirmed these preferences: Fos/Jun dimers enrich for TRE motifs with 7-bp spacing, while Jun/ATF combinations favor 8-bp spaced CRE variants, highlighting how dimer composition dictates motif selection.

Genome-Wide Target Identification

Genome-wide identification of AP-1 binding sites has primarily relied on followed by sequencing (ChIP-seq), which maps the locations of AP-1 subunits across the genome in various cell types and conditions. In stimulated mouse embryonic fibroblasts (MEFs), ChIP-seq analyses of AP-1 family members such as FOS, FOSL2, and JUND have identified approximately 55,919 binding sites, with over 90% located in promoter-distal regions including enhancers. These sites are enriched near active regulatory elements, such as those marked by H3K4me1 and accessible , highlighting AP-1's role in signal-dependent enhancer activation. Cell-type specificity is evident in immune cells, where (LPS) stimulation of bone marrow-derived macrophages (BMDMs) induces dynamic AP-1 binding. For instance, ChIP-seq for JunD in LPS-treated rat BMDMs revealed 18,124 binding peaks associated with 7,612 genes, predominantly at distal enhancers responsive to inflammatory signals. In mouse macrophages stimulated with Kdo2-lipid A (a TLR4 agonist mimicking LPS), over 10,000 sites were detected for subunits like ATF3, , and JunD, with additional inducible binding for Fos and JunB, totaling more than 50,000 unique AP-1 sites across family members. These findings underscore AP-1's context-dependent occupancy, with stimulation expanding binding to thousands of inflammatory response elements. In cancer cells, AP-1 binding exhibits similar scale but with distinct regulatory features. ChIP-seq in (TNBC) BT549 cells identified 11,670 Fra-1 sites and 14,201 c-Jun sites, with substantial overlap (84%) and enrichment at enhancers, including those within super-enhancers like the ZEB2 locus that drive invasiveness. Broader analyses in lung adenocarcinoma (A549) and (K562) cells confirm AP-1 hotspots—multi-subunit binding regions numbering 1,000 to 4,000—often overlapping super-enhancers, though only about 15% colocalize precisely, emphasizing AP-1's contribution to oncogenic enhancer networks. Advances since 2010 have integrated ChIP-seq with to link AP-1 binding to transcriptional output. In stimulated macrophages and fibroblasts, such integrations demonstrate that AP-1 occupies 20-30% of stimulus-inducible genes, particularly those involved in and responses, with binding correlating to dynamic changes. For example, in LPS-stimulated BMDMs, JunD ChIP-seq peaks overlap with differentially expressed genes, confirming direct regulation of and pathways. These multi-omics approaches reveal AP-1's selective activation of subsets of bound sites based on cellular context. Public databases like provide comprehensive AP-1 data, linking its binding to open regions across types. ChIP-seq datasets show AP-1 motifs enriched in accessible (e.g., DNase I hypersensitive sites) in over 100 lines, with AP-1 occupancy facilitating co-binding of other factors at enhancers. In oesophageal models using -referenced open profiles, AP-1 motifs appear in 65% of differentially accessible regions, associating with upregulated oncogenic genes. These resources enable cross- comparisons, revealing AP-1's conserved role in and gene regulation.

Functional Networks and Context-Dependence

AP-1 integrates into core regulatory networks that drive key cellular processes, particularly , , and . In networks, AP-1 activates genes such as cyclin D1 and c-Myc, which promote progression and oncogenic growth; for instance, AP-1 dimers bind to the cyclin D1 promoter in an AKT-dependent manner to enhance transcription during mitogenic signaling. Similarly, in inflammatory circuits, AP-1 upregulates COX-2 and IL-6, amplifying pro-inflammatory responses; AP-1 binds directly to the COX-2 promoter upstream of the transcription start site to induce expression in response to stimuli like TNFα. For , AP-1 targets matrix metalloproteinases (MMPs) and (VEGF), facilitating remodeling and ; AP-1 mediates VEGF-induced endothelial and by activating downstream transcriptional programs. These networks highlight AP-1's role as a hub coordinating multiple oncogenic and inflammatory pathways. The functional output of AP-1 is highly context-dependent, shaped by the specific dimer composition and co-regulators present in a given cellular . Different Jun/Fos heterodimers can either activate or repress target genes; for example, c-Jun/c-Fos dimers typically promote transcription, while JunD-containing complexes often exert repressive effects, such as in quiescent cells where JunD inhibits proliferation-associated genes. Co-regulators further modulate this, with factors like p300/CBP enhancing activation in proliferative contexts or HDACs promoting repression during stress; this dimer-specific and co-regulator-driven variability allows AP-1 to toggle between pro- and anti-tumorigenic roles depending on signals like MAPK pathway intensity. AP-1 participates in intricate feedback loops that fine-tune its own activity and amplify signaling cascades. It auto-regulates the jun and fos genes through direct binding to their promoters, creating positive feedback that sustains AP-1 levels during prolonged stimulation; the c-jun promoter, in particular, is positively autoregulated by Jun/AP-1 to reinforce immediate-early gene expression. Additionally, AP-1 engages in crosstalk with other transcription factors, such as STAT3, within inflammatory networks; in cytokine storms, AP-1 and STAT3 co-activate shared targets like IL-6, exacerbating systemic inflammation through a cooperative NF-κB-linked circuit. Recent single-cell RNA sequencing studies have illuminated AP-1's role as a central in tumor microenvironments, revealing heterogeneous activation patterns across types. Post-2020 analyses in clear renal carcinoma show AP-1 coordinating immune evasion and stromal remodeling at single-cell resolution, with elevated Jun/Fos activity in tumor-associated macrophages driving pro-metastatic inflammation. In , scRNA-seq identifies AP-1 as a regulator, where its inhibition disrupts tumor states and microenvironmental interactions, highlighting context-specific hubs that vary by tumor subtype and therapy response. These insights underscore AP-1's dynamic integration into multicellular networks, beyond bulk analyses.