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CENPA

Centromere protein A (CENP-A), encoded by the CENPA gene, is a histone H3 variant that serves as the epigenetic marker defining centromere identity on chromosomes.^[1] Unlike canonical histone H3, CENP-A replaces H3 within nucleosomes specifically at centromeric regions, forming specialized chromatin structures essential for kinetochore assembly and accurate chromosome segregation during cell division.^[2] This protein was first identified through autoantibodies in patients with CREST syndrome, a form of systemic sclerosis, highlighting its immunological relevance.^[1] CENP-A's structure features a conserved histone fold domain with over 60% sequence identity to histone H3 in its C-terminal region, enabling it to integrate into nucleosomes while altering their properties, such as increased flexibility in protruding DNA ends to facilitate protein recruitment.^[3] The protein consists of 140 amino acids, with a molecular weight of approximately 16 kDa, and includes a centromere-targeting domain (CATD) critical for its localization.^[3] Functionally, CENP-A nucleosomes recruit the constitutive centromere-associated network (CCAN) of proteins, forming the foundation for the inner kinetochore and ensuring proper microtubule attachment during mitosis.^[1] Assembly of CENP-A into centromeric chromatin is tightly regulated and occurs primarily in early G1 phase, independent of DNA replication, through a process mediated by the chaperone HJURP and the Mis18 complex.^[1] This deposition dilutes during S phase and is replenished to maintain centromere specificity across cell generations.^[1] Dysregulation of CENP-A, such as overexpression or ectopic incorporation outside centromeres, is implicated in genomic instability, contributing to aneuploidy in cancers like colorectal and hepatocellular carcinoma.^[1] Additionally, anti-CENP-A autoantibodies are biomarkers for autoimmune diseases, including scleroderma, where they correlate with chromosomal abnormalities.^[1]

Discovery and nomenclature

Discovery

CENPA, known as centromere protein A (CENP-A), was first identified in 1985 by William C. Earnshaw and Naomi Rothfield, who used autoimmune sera from patients with the CREST variant of scleroderma to detect centromere-specific antigens through immunofluorescence staining of human mitotic chromosomes. These sera revealed a family of proteins localized exclusively to the centromeric regions of metaphase chromosomes, with CENP-A appearing as a 17-kDa polypeptide among the initial three antigens (CENP-A, -B, and -C). This serological approach capitalized on the high specificity of autoantibodies in CREST patients, enabling the visualization and initial biochemical characterization of centromere components previously inaccessible by conventional methods. In the late 1980s and early 1990s, subsequent studies confirmed CENP-A's histone-like properties through biochemical fractionation and immunofluorescence techniques on human chromosomes. Earnshaw and colleagues demonstrated that CENP-A co-purifies with nucleosome core particles and histones upon extraction from HeLa cell nuclei, suggesting its integration into chromatin. Further purification and partial amino acid sequencing in 1991 by Palmer et al. established that CENP-A shares significant sequence similarity with histone H3, particularly in conserved regions critical for nucleosome assembly, marking it as the first identified centromere-specific histone variant. These findings were supported by immunofluorescence assays showing CENP-A's exclusive localization to inner kinetochore regions during mitosis, distinguishing it from canonical histones. The cloning of the human CENPA gene occurred in the mid-1990s, providing definitive molecular evidence of its histone H3 variant nature and centromere-specific targeting. In 1994, Sullivan et al. isolated a full-length cDNA encoding the 140-amino-acid CENP-A protein, revealing a histone fold domain homologous to H3 but with unique N-terminal extensions essential for centromeric localization. Transfection experiments with epitope-tagged CENP-A constructs confirmed its specific accumulation at centromeres in mitotic cells, independent of DNA sequence, through immunofluorescence and subcellular fractionation. These key experiments solidified CENP-A's role as a structural component of centromeric chromatin. By the 2000s, research milestones highlighted CENP-A as a pivotal epigenetic marker for centromere identity and inheritance. Studies, such as those by Blower et al. in 2002, showed that CENP-A nucleosomes are stably propagated through cell divisions, serving as a heritable mark that defines active centromeres without relying on underlying DNA sequences. This recognition culminated in experiments demonstrating that ectopic deposition of CENP-A could induce neocentromere formation, underscoring its sufficiency as an epigenetic specifier in human cells.00664-3)

Nomenclature and gene location

The CENPA gene, officially designated as centromere protein A by the HUGO Gene Nomenclature Committee (HGNC), carries the approved symbol CENPA (HGNC ID: 1851).^[4] It was previously referred to as CENP-A, a nomenclature reflecting its identification as a centromeric antigen.^[5] In humans, the CENPA gene is located on the short arm of chromosome 2 at cytogenetic band 2p23.3, with genomic coordinates spanning from 26,764,289 to 26,801,067 on the forward strand (GRCh38.p14 assembly).^[5] The gene covers approximately 37 kb and consists of 5 exons.^[5] CENPA exhibits strong evolutionary conservation as a histone H3 variant essential for centromere function, with orthologs including Cid (centromere identifier) in Drosophila melanogaster and Cse4 in budding yeast (Saccharomyces cerevisiae).^[6] No pseudogenes for CENPA have been identified in the human genome, and the locus resides in a region flanked by other protein-coding genes on chromosome 2, such as those involved in cellular processes unrelated to centromere biology.^[3]

Structure

Gene structure

The CENPA gene spans approximately 8,534 base pairs on the short arm of human chromosome 2 at locus 2p23.3 and consists of five exons interrupted by four introns, with the primary transcript encoding a 140-amino acid protein.^[5] The first exon includes non-coding regions encompassing the promoter, while the coding sequence is distributed across the exons to produce the mature mRNA.^[7] The promoter of CENPA is TATA-less and GC-rich, featuring multiple binding sites for the transcription factor Sp1, which facilitates basal and cell cycle-dependent transcriptional activation to synchronize expression with DNA replication and mitosis.^[3]^[8] CENPA mRNA is expressed at low ubiquitous levels across human tissues, such as lymph nodes and appendix, but is upregulated in proliferating cells to support kinetochore assembly during active cell division.^[5] Alternative splicing events are infrequent and yield minor isoforms with limited functional divergence from the canonical form.^[5]^[9] The genomic organization and sequence of CENPA are highly conserved among eukaryotes, reflecting its essential role in centromere function; the human protein shares 60-70% sequence identity with the Saccharomyces cerevisiae homolog Cse4, particularly within the core histone fold domain.^[10]^[11]

Protein structure

The human CENP-A protein is a 140-amino-acid histone H3 variant with a molecular weight of approximately 16 kDa.^[10] Like canonical histone H3, it possesses a central histone fold domain (HFD) composed of three α-helices (α1, α2, and α3) flanked by shorter loops, enabling incorporation into nucleosomes.^[12] However, CENP-A diverges from H3 in its N-terminal tail, which spans the first ~40 residues and is enriched in serine and threonine residues that serve as sites for post-translational modifications such as phosphorylation. This tail precedes a shorter αN helix, consisting of only two helical turns compared to three in H3, which influences the overall nucleosomal architecture.^[13] A defining feature of CENP-A is the centromere targeting domain (CATD), embedded within the HFD and encompassing loop 1 and the α2 helix. The CATD confers specificity for centromeric chromatin by forming a rigid, hydrophobic interface that distinguishes CENP-A from H3.^[14] Structural analyses reveal that the CATD induces a rotated orientation in the CENP-A–CENP-A dimer interface relative to H3–H3, along with an oppositely charged loop 1 region, resulting in a more compact (CENP-A/H4)₂ tetramer.^[14] These alterations enhance the rigidity of the histone core while reducing interactions at the nucleosome periphery. Key differences from H3 include the shortened αN helix and the rigid CATD, which collectively destabilize DNA wrapping at the nucleosome entry and exit sites.^[15] In canonical H3 nucleosomes, 147 base pairs of DNA are tightly wrapped in 1.65 left-handed superhelical turns; in contrast, CENP-A nucleosomes stably protect only about 121 base pairs, with the terminal ~13 base pairs at each end exhibiting greater flexibility and accessibility.^[13] This unwrapping promotes an open chromatin conformation at centromeres. High-resolution crystal structures have elucidated these features. The (CENP-A/H4)₂ tetramer structure (PDB: 3NQJ) highlights the compact, disk-shaped assembly and rotated interfaces that mark centromeric identity from within the histone core.^[14] The full CENP-A nucleosome structure (PDB: 3AN2), solved with α-satellite DNA, confirms the octameric composition and loose DNA ends, underscoring how CENP-A remodels nucleosome geometry for specialized function.^[13]

Biological function

Centromere specification

CENP-A nucleosomes serve as the primary epigenetic mark that defines and maintains centromere identity by replacing canonical histone H3 nucleosomes within centromeric chromatin.^[16] In humans, this occurs predominantly at alpha-satellite DNA, which consists of tandem 171-bp repeat units forming higher-order arrays that span megabases of pericentromeric heterochromatin.^[17] The incorporation of CENP-A imparts structural rigidity to these nucleosomes, distinguishing them from H3-containing ones and enabling heritable propagation of centromeric specification independent of underlying DNA sequence.^[16] The assembly pathway for centromeric chromatin involves the targeted replacement of H3 nucleosomes with CENP-A-H4 tetramers, which form the foundational layer of the Constitutive Centromere Associated Network (CCAN).^[18] This process relies on the CENP-A targeting domain (CATD), a region homologous to H3's alpha-2 helix, which directs the stable deposition of CENP-A into octameric nucleosomes alongside histones H2A, H2B, and H4.^[18] These CENP-A nucleosomes then recruit CCAN components, such as CENP-C and CENP-N, to establish a self-propagating chromatin domain that perpetuates centromere function across cell divisions.^[18] Evidence for the epigenetic basis of centromere specification is provided by neocentromeres in humans, where CENP-A marks functional centromeric sites that form de novo on non-alpha-satellite DNA sequences.^[19] Over 100 such cases have been documented, involving derivatives of nearly all human chromosomes, including examples like the mardel(10) marker chromosome at 10q25, which lacks alpha-satellite but binds CENP-A and supports stable kinetochore assembly and chromosome segregation.^[20] These neocentromeres demonstrate that CENP-A deposition can initiate and maintain centromeric identity in ectopic locations, often associated with chromosomal rearrangements in disease contexts.^[19] CENP-A nucleosome stability is maintained through a balance of dilution during S phase DNA replication and replenishment in early G1 phase, ensuring consistent centromeric marking.^[21] During replication, existing CENP-A nucleosomes are randomly segregated to daughter strands, halving their density, after which new CENP-A is loaded to restore levels.^[21] This results in a steady-state density of approximately one CENP-A nucleosome per 5-10 kb across the centromeric domain, with human centromeres typically harboring around 200 such nucleosomes within a centromeric domain of approximately 1 Mb.^[21]

Kinetochore assembly

CENP-A nucleosomes form the foundational platform for kinetochore assembly by directly recruiting key components of the constitutive centromere-associated network (CCAN), particularly CENP-C and CENP-N. The C-terminal domain of CENP-C binds to the RG loop and CATD region of the CENP-A/H4 dimer within the nucleosome, stabilizing its conformation and enabling further CCAN recruitment. Similarly, CENP-N recognizes the L1 loop of CENP-A and interacts with approximately 15 base pairs of distorted nucleosomal DNA through charge and space complementarity, involving residues such as E3, E7, R11, K143, P145, N146, and K148. These bindings initiate a hierarchical assembly process, where CENP-C and the CENP-L/N complex bridge to the outer kinetochore KMN network—comprising KNL1, Mis12, and Ndc80 complexes—to facilitate microtubule attachment.^[22]^[23]^[24] Through its integration into the CCAN, CENP-A contributes to the formation of the inner kinetochore layer, which provides a robust structural scaffold at the centromere. Cryo-electron microscopy structures reveal that the 16-subunit CCAN complex encircles linker DNA emerging from CENP-A nucleosomes in an edge-on manner, with the CENP-L/N and CENP-T/WSX modules gripping and partially wrapping the DNA to create topological entrapment. This configuration ensures stability against mitotic forces, allowing the inner kinetochore to serve as a pedestal for outer kinetochore components, including the KMN network and microtubule-binding proteins like the Ndc80 complex, thereby enabling bi-orientation and attachment to spindle microtubules.^[25]^[26] The essential role of CENP-A in kinetochore assembly is evident from depletion studies, which demonstrate its necessity for mitotic fidelity. In human and chicken DT40 cells, RNAi-mediated CENP-A depletion impairs CCAN recruitment, leading to defective kinetochore formation, chromosome misalignment on the metaphase plate, and extended prometaphase arrest—such as a 12-fold increase in prometaphase cells. Consequently, chromosome segregation fails, resulting in lagging chromosomes, unequal DNA distribution in 92% of anaphases, increased multinucleation, and aneuploidy in daughter cells.^[27] CENP-A's function in kinetochore assembly is evolutionarily conserved across eukaryotes, from the yeast homolog Cse4, which incorporates into a single or limited number of nucleosomes at short point centromeres (~125 bp), to humans, where arrays of multiple CENP-A nucleosomes span megabase-scale regional centromeres defined by α-satellite DNA. Despite these structural differences—point centromeres in yeast relying on sequence-specific factors like CBF3 versus epigenetic specification in humans—the core mechanism of CENP-A nucleosomes recruiting CCAN and outer kinetochore components remains invariant, highlighting its pivotal role in chromosome segregation.^[28]^[29]

Regulation

Chaperone-mediated deposition

The chaperone-mediated deposition of CENPA (centromere protein A) at centromeres involves a coordinated process that ensures the specific replacement of canonical histone H3 nucleosomes with CENPA-containing nucleosomes. Central to this is the Mis18 complex, composed of Mis18α, Mis18β, and M18BP1 (also known as KNL2), which acts post-mitosis to create a receptive chromatin state at centromeres. This complex localizes to centromeres during late mitosis and early G1 phase, where it promotes the destabilization and eviction of H3 nucleosomes, thereby priming the site for new CENPA loading.^[30]^[31] The primary chaperone for CENPA is HJURP (Holliday junction recognition protein), which binds the newly synthesized CENPA-H4 histone tetramer and delivers it to the centromeric DNA. HJURP recognizes CENPA through its centromere targeting domain (CATD), a conserved region in the histone fold that distinguishes CENPA from H3 and facilitates specific chaperone binding. This interaction enables HJURP to target alpha-satellite DNA sequences characteristic of human centromeres, where it deposits the CENPA-H4 tetramer onto the DNA, forming stable nucleosomes essential for centromere identity. The Mis18 complex recruits HJURP to these sites, ensuring deposition occurs only at appropriate locations and times.^[32]^[33] Experimental evidence for this mechanism comes from in vitro reconstitution assays using purified recombinant proteins, which demonstrate that HJURP is both necessary and sufficient for assembling CENPA-containing nucleosomes on DNA templates. In these assays, HJURP facilitates the efficient deposition of CENPA-H4 tetramers onto naked DNA, mimicking centromeric chromatin, while omission of HJURP abolishes assembly. Further studies confirm that the Mis18 complex enhances this process by preparing the chromatin substrate, underscoring the interdependence of these factors in vivo.^[34]^[32]^[33]

Cell cycle control

The expression of CENPA (also known as CENP-A) is tightly regulated throughout the cell cycle to ensure its precise incorporation into centromeric chromatin. Transcription of the CENPA gene primarily occurs during the G2 phase in human cells, allowing for the accumulation of mRNA that peaks during mitosis. This timing decouples CENPA synthesis from DNA replication, preventing interference with S-phase processes. The resulting protein is then available for chaperone-mediated deposition, such as by HJURP, specifically in late mitosis (telophase) and extending into early G1 phase. Additionally, polo-like kinase 1 (PLK1) phosphorylates M18BP1 at threonine 78 and serine 93, and Mis18α at serine 54 during G1, licensing the Mis18 complex to recruit HJURP and promote CENPA loading.^[35]^[36]^[37]^[38]^[39] During S phase, existing CENP-A nucleosomes at centromeres undergo passive dilution as DNA replication proceeds, with each parental nucleosome randomly distributed to one of the two daughter strands, effectively halving the CENP-A content per centromere. This dilution mechanism maintains centromere identity without over-accumulation, as new CENP-A is replenished only in the subsequent G1 phase to restore full levels. The restriction of deposition to post-replicative G1 avoids conflicts with replication forks and ensures stable propagation of centromeric chromatin across divisions.^[40]^[41]^[42] CENP-A regulation integrates with cell cycle checkpoints, particularly those involving the DNA damage response (DDR). Upon detection of double-strand breaks, CENP-A is rapidly recruited to damage sites alongside DDR components like CENP-N, facilitating repair while preserving centromeric function. Additionally, during early mitosis, CDK1 phosphorylates CENP-A at serine 68, which disrupts its interaction with the chaperone HJURP and inhibits premature deposition until mitotic exit. This phosphorylation event ensures that new CENP-A loading aligns with the completion of chromosome segregation.^[43]^[44] Dysregulation of this temporal control, such as mistimed deposition outside of G1, can lead to ectopic incorporation of CENP-A at non-centromeric sites. This misplacement disrupts normal chromatin architecture and kinetochore assembly, resulting in chromosomal instability through mechanisms like anaphase bridges and lagging chromosomes. Such errors compromise the fidelity of segregation, highlighting the critical need for phase-specific barriers in CENP-A handling.^[45]^[46]

Interactions

Key protein partners

CENPA, the histone H3 variant essential for centromere identity, forms stable interactions with several core protein partners within the constitutive centromere-associated network (CCAN). Among these, CENPC directly binds CENPA nucleosomes via the RG loop in the L1 region of CENPA, enabling specific recognition and stabilization of centromeric chromatin.^[47] This interaction is mediated by the C-terminal domain of CENPC, which engages hydrophobic and electrostatic contacts with the CENPA-specific features.^[47] Similarly, CENPN recognizes the faces of the CENPA nucleosome, primarily through electrostatic and hydrophobic interactions with the L1 loop (also known as the RG loop), conferring binding specificity over canonical H3 nucleosomes.^[48] HJURP, the dedicated chaperone for CENPA, transiently associates with CENPA/H4 dimers via its N-terminal CENPA-binding domain to facilitate nucleosome loading, but dissociates after deposition.^[32] Beyond these core interactors, CENPA is embedded in a broader CCAN network comprising associations with multiple subunits, including the CENPL/CENPM heterodimer and the CENPH/I/K complex. The CENPL-N complex serves as a central node, bridging CENPA-bound CENPC and the CENPH-I-K-M module to form an integrated 16-subunit assembly that maintains centromere integrity.^[49] These interactions are interdependent, with CENPC recruiting CENPL-N and stabilizing its localization, while pairwise contacts between CENPL-N and CENPH-I-K-M ensure the structural cohesion of the CCAN.^[49] Quantitative assessments of binding affinities, such as those from pulldown assays, reveal a high-affinity interaction between CENPA nucleosomes and CENPC, underscoring the robustness of this partnership.^[50] Competitive pulldowns further demonstrate that phosphorylated CENPC efficiently displaces other binders like CENPN from the CENPA nucleosome, highlighting the selective nature of these associations.^[50] CENPA's partnerships exhibit dynamic exchanges across the cell cycle, reflecting regulatory swaps to coordinate centromere function. For instance, HJURP dissociates from CENPA post-deposition in early G1, allowing CENPC to engage stably during subsequent phases, a transition enforced by phosphorylation events that inhibit HJURP-CENPC interactions in metaphase.^[51] This timing aligns with chaperone-mediated deposition primarily in G1, ensuring precise control over CENPA incorporation.^[51]

Post-translational modifications

CENPA, the centromeric histone H3 variant, undergoes several post-translational modifications (PTMs) that regulate its stability, localization to centromeres, and eviction from chromatin. These modifications include phosphorylation, acetylation, ubiquitination, and sumoylation, primarily identified through mass spectrometry analyses of human and yeast orthologs. Mass spectrometry studies have mapped approximately 10 modifiable residues on CENPA, with many conserved across species, such as serines in the N-terminal tail and lysine 124 in the histone fold domain. For instance, high-resolution mass spectrometry has detected trimethylation at the α-amino group of Gly1, phosphorylation at Ser16 and Ser18 in the N-tail, and acetylation at Lys124, highlighting the dynamic PTM landscape that distinguishes CENPA from canonical H3.^[52]^[53] Phosphorylation is a prominent PTM on CENPA, particularly at Ser7 in the N-terminal tail, catalyzed by Aurora kinases (A and B) during prophase and mitosis for involvement in kinetochore assembly and mitotic progression.^[52]^[54] This modification peaks in prometaphase and declines in anaphase. Multiple serine phosphorylations in the N-tail, including Ser16 and Ser18, further influence CENPA solubility and eviction from non-centromeric sites; double phosphorylation at these residues forms a salt-bridged structure that compacts the tail, reducing nucleosome oligomerization and enhancing solubility in prenucleosomal forms. These N-tail dynamics ensure precise centromere maintenance by preventing ectopic accumulation, with phospho-mimetic mutants showing altered chromatin compaction similar to canonical H3 arrays. Additionally, phosphorylation at Ser68 primes ubiquitination-dependent degradation during mitosis, linking PTMs in a coordinated regulatory cascade.^[52]^[54]^[53]^[55] Acetylation at Lys124, mediated by the histone acetyltransferase p300 during G1/S phase, plays a key role in chaperone release and nucleosome dynamics. This modification tightens the histone core structure, hindering accessibility to the C-terminus and potentially impeding recruitment of kinetochore proteins like CENP-C, while facilitating the dissociation from the chaperone HJURP to enable deposition. Computational modeling and in vivo assays confirm that Lys124 acetylation loosens DNA wrapping at the nucleosome dyad, promoting asymmetric unwrapping and replication timing shifts from early to mid-S phase. Ubiquitination, often at Lys124 by the CUL4A-RBX1-COPS8 E3 ligase complex, supports deposition by enhancing HJURP binding but also marks CENPA for proteasomal degradation when primed by Ser68 phosphorylation, preventing excess accumulation and ensuring cell-cycle-timed turnover.^[56]^[52]^[57]^[55] Sumoylation of the yeast ortholog Cse4, with conserved mechanisms affecting human CENPA stability, contributes to targeting the variant for degradation via ubiquitin ligases like Slx5-Slx8, thereby restricting ectopic incorporation and maintaining epigenetic boundaries. Overall, these PTMs collectively fine-tune CENPA's chromatin association, with phosphorylation driving eviction and acetylation/ubiquitination balancing deposition and degradation for faithful centromere inheritance. Recent studies (as of 2024) highlight SUMO pathway involvement in CENPA turnover via p97/VCP and chaperones like DNAJC9 preventing mislocalization.^[52]^[58]^[59]

Clinical significance

Role in cancer

CENPA is overexpressed in a wide range of solid tumors, including breast, lung, colorectal, prostate, and ovarian cancers, where elevated levels correlate with advanced tumor grade, increased invasiveness, and poor patient prognosis.^[60] In many cancer types, CENPA mRNA and protein expression exceed normal levels by up to 1000-fold compared to healthy tissues, contributing to disease progression across approximately 20 distinct malignancies.^[61] For instance, in breast cancer, high CENPA expression is associated with higher risk of relapse and metastasis, particularly in node-negative, estrogen receptor-positive (luminal A) subtypes, serving as an independent prognostic marker.^[62] A genotranscriptomic meta-analysis across multiple cancer datasets further confirms that CENPA upregulation occurs in many analyzed tumors and predicts adverse outcomes, including reduced overall survival.^[63] The oncogenic mechanisms driven by CENPA overexpression primarily involve its ectopic deposition outside centromeres, forming non-centromeric nucleosomes that disrupt chromatin architecture and promote chromosomal instability (CIN). Mislocalized CENPA nucleosomes often assemble at gene promoters, including those of oncogenes, leading to aberrant gene expression, mitotic errors, and aneuploidy that fuel tumorigenesis.^[60] This mislocalization is exacerbated in cancer cells with p53 deficiencies, where excess CENPA triggers epithelial-to-mesenchymal transition (EMT) and enhances metastatic potential, while in p53-wild-type cells, it induces cell cycle arrest but still contributes to genomic instability over time.^[60] In breast cancer models, CENPA-driven CIN correlates with tumor heterogeneity and resistance to therapy, underscoring its role in sustaining aneuploidy as a cancer hallmark.^[46] Therapeutically, CENPA serves as a promising biomarker for prognosis and treatment stratification, with high levels indicating susceptibility to interventions targeting its deposition pathway. Inhibitors of the CENPA chaperone HJURP, which facilitate its centromeric loading, have shown potential to reduce ectopic assembly and impair cancer cell proliferation in preclinical models of ovarian and breast cancers.^[64] Similarly, Aurora B kinase inhibitors, which regulate CENPA phosphorylation and kinetochore function, disrupt mitotic fidelity in CENPA-overexpressing tumors, enhancing sensitivity to chemotherapy and radiation in solid malignancies like lung and colorectal cancers.^[65] These approaches exploit cancer-specific vulnerabilities in centromere maintenance, with ongoing research exploring CENPA-targeted therapies to mitigate CIN without affecting normal cells.^[66]

Association with other diseases

CENPA serves as an autoantigen in autoimmune disorders, particularly in limited cutaneous systemic sclerosis (lcSSc), also known as CREST syndrome, where anti-centromere antibodies (ACA) targeting CENPA and other centromeric proteins are detected in 20-40% of patients.^[67] These antibodies are more prevalent in the limited form of the disease compared to diffuse systemic sclerosis, contributing to diagnostic specificity for vascular and fibrotic manifestations.^[68] In developmental disorders, CENPA function is indirectly disrupted in immunodeficiency, centromere instability, and facial anomalies (ICF) syndrome, an autosomal recessive condition caused by mutations in genes such as DNMT3B, which interacts with centromeric proteins to regulate DNA methylation at centromeres.^[69] Hypomethylation of pericentromeric regions in ICF leads to centromere instability and chromosomal breakage, particularly at heterochromatic regions.^[70] CENPA expression declines with age in human pancreatic β-islet cells, dropping sharply by around 30 years and contributing to reduced cellular proliferation and adaptive responses to metabolic stress.^[71] This age-related reduction is associated with chromosome instability in aging tissues, including potential links to neurodegeneration; for instance, centromere instability on the X chromosome has been observed in Alzheimer's disease brains, suggesting a broader role for centromeric defects in neuronal vulnerability.^[72]

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Point centromeres contain more than a single centromere-specific ...
Nov 14, 2011 · Cse4 is the budding yeast homologue of CENP-A, a modified histone H3 that specifies the base of kinetochores in all eukaryotes. Budding ...Missing: similarity | Show results with:similarity
[29]
Centromere DNA Destabilizes H3 Nucleosomes to Promote CENP ...
Nov 29, 2018 · Analysis of human centromere chromatin fibers suggested that H3.3 is deposited as a placeholder in S phase that is later replaced by new CENP-A ...
[30]
Mechanism of centromere recruitment of the CENP-A chaperone ...
Sep 6, 2019 · HJURP is the CENP-A chaperone, which associates with Mis18α, Mis18β, and M18BP1 to target centromeres and deposit new CENP-A.
[31]
HJURP binds CENP-A via a highly conserved N-terminal domain ...
We found that HJURP, a member of the complex, was required for cell cycle specific targeting of CENP-A to centromeres. HJURP facilitated efficient deposition of ...
[32]
HJURP is a CENP-A chromatin assembly factor sufficient to form a ...
Jul 18, 2011 · Together, these data suggest that the role of the Mis18 complex in CENP-A deposition is to recruit HJURP and that the CENP-A nucleosome assembly ...
[33]
HJURP is a CENP-A chromatin assembly factor sufficient to form a ...
We assessed the ability of HJURP to assemble CENP-A–containing nucleosomes in an in vitro chromatin assembly assay using purified recombinant proteins ...
[34]
Assembly of CENP-A into centromeric chromatin requires ... - PubMed
Feb 10, 1997 · We investigated the requirements for targeting the centromeric histone H3 homologue CENP-A for assembly at centromeres in human cells.
[35]
HJURP Is a Cell-Cycle-Dependent Maintenance and Deposition ...
May 1, 2009 · HJURP downregulation leads to a major reduction in CENP-A at centromeres and impairs deposition of newly synthesized CENP-A, causing mitotic defects.
[36]
Propagation of centromeric chromatin requires exit from mitosis
Mar 5, 2007 · The CENP-A–SNAP pool produced during mid-to-late G1 was labeled and monitored for timing of centromeric deposition (Fig. 4). DNA content was ...
[37]
CENP-A nucleosome clusters form rosette-like structures around ...
Sep 30, 2019 · Here we show that CENP-A nucleosomes form characteristic clusters during the G1 phase of the cell cycle. 2D and 3D super-resolution microscopy ...
[38]
Inheritance of the CENP-A chromatin domain is spatially and ...
May 31, 2016 · We demonstrate that most human chromosomes share similar patterns of CENP-A loading and distribution and that centromere inheritance is achieved through ...
[39]
Assembly in G1 phase and long-term stability are unique intrinsic ...
Jan 30, 2013 · We previously showed that G1 phase–restricted assembly of CENP-A in human and chicken cells is directly coupled to cell cycle progression as a ...
[40]
Double-strand DNA breaks recruit the centromeric histone CENP-A
Abstract. The histone H3 variant CENP-A is required for epigenetic specification of centromere identity through a loading mechanism independent of DNA sequence.
[41]
Dynamic Phosphorylation of CENP-A at Ser68 Orchestrates Its Cell ...
Jan 12, 2015 · Here, we show that CENP-A is phosphorylated at Ser68 during early mitosis by Cdk1. Our results demonstrate that phosphorylation of Ser68 ...
[42]
Mislocalization of centromeric histone H3 variant CENP-A ...
Jul 18, 2017 · Our results show that mislocalization of overexpressed CENP-A to chromosome arms leads to chromosome congression defects, lagging chromosomes, micronuclei ...
[43]
CENP-A overexpression promotes aneuploidy with karyotypic ...
Feb 23, 2021 · ... Cse4 in budding yeast, Cid in flies—at α satellite DNA motifs ... DNA replication also acts as a mechanism for maintaining centromere identity ...
[44]
Cryo‐EM structure of the CENP‐A nucleosome in complex with ...
Jan 19, 2021 · Cryo‐EM reveals how CENP‐C phosphorylation regulates CENP‐A binding and provides insights into a dynamic kinetochore assembly mechanism during mitosis.Missing: pulldown | Show results with:pulldown
[45]
Structural mechanisms of centromeric nucleosome recognition by ...
We show that human CENP-N confers binding specificity through interactions with the L1 loop of CENP-A, stabilized by electrostatic interactions with the ...Missing: partners components
[46]
The CENP-L-N complex forms a critical node in an integrated ...
We demonstrate that the CENP-LN complex plays a crucial role at the core of this assembly through interactions with CENP-C and CENP-HIKM.
[47]
Cryo‐EM structure of the CENP‐A nucleosome in complex with phosphorylated CENP‐C | The EMBO Journal
### Summary of CENP-C Binding to CENP-A RG Loop
[48]
Repression of CENP-A assembly in metaphase requires HJURP ...
We have previously shown that changes in CENP-A levels in this assay represent new assembly and not simply tethering of CENP-A through HJURP (Westhorpe et al.
[49]
Posttranslational modifications of CENP-A: marks of distinction - PMC
Mar 22, 2018 · CENP-A undergoes posttranslational modifications including phosphorylation, acetylation, methylation, and ubiquitylation.
[50]
Posttranslational modification of CENP-A influences the ... - PNAS
Jul 1, 2013 · We report three posttranslational modifications on human CENP-A N termini using high-resolution MS: trimethylation of Gly1 and phosphorylation of Ser16 and Ser ...
[51]
Phosphorylation of CENP-A on serine 7 does not control centromere ...
Jan 11, 2019 · The amino-terminal tail of CENP-A also takes part in CENP-C mobilization but indirectly via its interaction with the DNA binding protein CENP-B ...
[52]
Phosphorylation at Ser68 facilitates DCAF11-mediated ...
Nov 9, 2021 · We report that the phosphorylation of CENP-A Ser68 primes the ubiquitin-proteasome-mediated proteolysis of CENP-A during mitotic phase in human cultured cells.
[53]
Internal modifications in the CENP-A nucleosome modulate ...
Apr 4, 2017 · Our data suggest that cyclical modifications within the CENP-A nucleosome contribute to the binding of key kinetochore proteins, the integrity of mitosis, and ...Cenp-A K124 Switches From... · Hats And Hdacs Influencing... · Additional Files
[54]
CENP-A K124 Ubiquitylation Is Required for CENP-A Deposition at ...
CENP-A is a centromere-specific histone H3 variant that epigenetically determines centromere identity to ensure kinetochore assembly and proper chromo-some ...
[55]
CENP-A overexpression promotes distinct fates in human cells ...
Mar 26, 2021 · CENP-A, the centromeric histone H3 variant, is an epigenetic mark that directly perturbs genetic stability and chromatin when overexpressed.
[56]
Guarding the Genome: CENP-A-Chromatin in Health and Cancer
... identity [8,17], as evidenced by the growing numbers of identified ... (Cenpa) null mice. Proc. Natl. Acad. Sci. USA. 2000;97:1148–1153. doi: 10.1073 ...
[57]
Centromere protein-A, an essential ... - Breast Cancer Research
May 4, 2012 · We discovered that CENP-A is an independent prognostic marker for relapse in node-negative, ER-positive breast cancers not treated with systemic ...Patient Populations · Gene Expression Analysis · Cenp-A And Cenp-B Levels
[58]
Genotranscriptomic meta-analysis of centromere protein A (CENPA ...
Aug 1, 2015 · Conclusions: The current study highly indicates the essential cancer promoting role of CENPA in human cancer and its value as prognostic marker ...
[59]
Advances in holliday junction recognition protein (HJURP) - Frontiers
The localization of HJURP at centromeres is cell-cycle regulated, and CENP-A expression is reduced after inhibition of HJURP (Zasadzińska et al., 2013). Upon ...Abstract · Introduction · Molecular functions of HJURP · Roles of HJURP in tumors
[60]
Aurora B promotes the CENP-T–CENP-W interaction to guide ...
Aurora B promotes the CENP-T–CENP-W interaction to guide accurate chromosome segregation in mitosis.
[61]
m6A-modified cenRNA stabilizes CENPA to ensure centromere ...
Oct 17, 2024 · The m 6 A-modified cenRNA stabilizes centromeric localization of CENPA in cancer cells during the S phase of the cell cycle.Missing: meta- prognosis
[62]
Clinical and serological evaluation of a novel CENP-A peptide ...
May 20, 2010 · Anti-centromere antibodies (ACA) are useful biomarkers in the diagnosis of systemic sclerosis (SSc). ACA are found in 20 to 40% of SSc patients ...
[63]
Anti-centromere antibodies (ACA) | Clinical Rheumatology
ACA are clinically associated with the CREST syndrome or limited cutaneous systemic sclerosis (prevalence ±55%), and are also found in 30% of patients with ...
[64]
DNMT3B interacts with constitutive centromere protein CENP-C to ...
DNA methyltransferase 3B (DNMT3B) mutations in ICF syndrome lead to altered epigenetic modifications and aberrant expression of genes regulating development ...
[65]
DNA methylation influences human centromere positioning ... - Nature
Sep 4, 2025 · The increase in CENP-A levels could reflect a higher density of CENP-A nucleosomes within the same region or, alternatively, could mean that ...
[66]
Assessment of X Chromosome Centromere Instability in Alzheimer's ...
Jun 5, 2025 · This pilot study aimed to establish a quantitative technique for assessing X chromosome centromere signals using fluorescence in situ hybridization (FISH).