Affinage

CENPH

Centromere protein H · UniProt Q9H3R5

Length
247 aa
Mass
28.5 kDa
Annotated
2026-06-09
16 papers in source corpus 12 papers cited in narrative 12 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CENP-H is a constitutive inner kinetochore protein that localizes to centromeres throughout the cell cycle and is required for proper kinetochore assembly and chromosome segregation (PMID:10488063, PMID:11500386). Within the centromere assembly hierarchy, CENP-A localization is upstream of CENP-H, and CENP-H is in turn required to recruit CENP-C—but not CENP-A—to the centromere, with its loss producing centromere dysfunction and metaphase arrest (PMID:11500386); CENP-H and CENP-C co-occupy discontinuous CENP-A chromatin subdomains at neocentromeres, defining the inner kinetochore chromatin architecture (PMID:17651496). CENP-H forms a stable, salt-resistant ~1:1 coiled-coil heterodimer with CENP-K and connects the inner kinetochore to the outer kinetochore through interaction with Hec1 of the Nuf2 complex (PMID:15713649, PMID:19381461). Functionally, CENP-H cooperates with CSPP1 to regulate kinetochore–microtubule dynamics, chromosome oscillation, and spindle assembly checkpoint satisfaction (PMID:26378239). CENP-H is essential for mitosis in vivo: its loss in zebrafish causes chromosome missegregation, G2/M arrest, and p53-dependent intrinsic apoptosis, and CENPH heterozygosity reduces tumor invasion (PMID:20573960). In meiosis, CENP-H is required for the oocyte G2/M transition by protecting cyclin B1 from APC/C(Cdh1)-mediated degradation, thereby enabling MPF activation (PMID:27993978). Beyond its kinetochore role, CENP-H physically interacts with GOLPH3 to attenuate mTORC1/mTORC2 signaling (PMID:28819418).

Mechanistic history

Synthesis pass · year-by-year structured walk · 10 steps
  1. 1999 Medium

    Established CENP-H as a bona fide constitutive kinetochore component, defining a structural protein present throughout the cell cycle rather than a transiently recruited factor.

    Evidence Protein isolation, sequence analysis, and immunofluorescence with anti-CENP-H antibody

    PMID:10488063

    Open questions at the time
    • Did not define functional partners or the assembly hierarchy
    • Coiled-coil and NLS inferred from sequence, not functionally tested
  2. 2001 High

    Positioned CENP-H within the centromere assembly hierarchy by showing it is required to recruit CENP-C but not CENP-A, establishing CENP-A as the upstream determinant.

    Evidence Conditional loss-of-function knockout in chicken DT40 cells with immunocytochemistry

    PMID:11500386

    Open questions at the time
    • Molecular basis of CENP-C recruitment not defined
    • Direct vs indirect dependence not resolved
  3. 2005 High

    Identified the physical bridge between inner and outer kinetochore by showing CENP-H stably associates with Hec1 of the Nuf2 complex.

    Evidence Yeast two-hybrid, reciprocal co-immunoprecipitation, and FRAP in chicken DT40 cells

    PMID:15713649

    Open questions at the time
    • Stoichiometry and structural interface with Hec1 unresolved
    • Functional consequence of disrupting the interaction not tested
  4. 2006 Medium

    Extended CENP-H's architectural role to human cells, showing it supports CENP-E recruitment and chromosome alignment without being strictly required for mitotic arrest.

    Evidence RNAi knockdown with immunofluorescence and Western blot in human HEp-2 cells

    PMID:16875666

    Open questions at the time
    • Mechanism of CENP-E recruitment not defined
    • Partial CENP-C reduction not mechanistically explained
  5. 2007 Medium

    Defined the inner kinetochore chromatin organization by mapping CENP-H and CENP-C to discontinuous CENP-A subdomains, supporting higher-order chromatin looping.

    Evidence ChIP-on-chip on neocentromeres using BAC and PCR-amplicon microarrays

    PMID:17651496

    Open questions at the time
    • Looping model not directly demonstrated
    • Restricted to neocentromere context
  6. 2009 Medium

    Resolved a core subcomplex by demonstrating CENP-H and CENP-K form a stable ~1:1 coiled-coil heterodimer within the inner kinetochore.

    Evidence Tandem affinity purification from HEK293 cells with bioinformatic coiled-coil domain mapping

    PMID:19381461

    Open questions at the time
    • No high-resolution structure of the heterodimer
    • Functional validation of mapped interaction regions lacking
  7. 2010 High

    Established CENP-H as essential for mitosis in vivo and linked it to tumor suppression, connecting kinetochore integrity to organismal viability and cancer.

    Evidence Zebrafish transposon mutant, morpholino, mRNA rescue, p53 epistasis, and in vivo tumor incidence assay

    PMID:20573960

    Open questions at the time
    • Molecular trigger of intrinsic apoptosis downstream of missegregation not defined
    • Mechanism of tumor suppression beyond mitotic fidelity unclear
  8. 2015 High

    Identified CSPP1 as a CENP-H-interacting regulator of kinetochore–microtubule dynamics and showed the interaction is functionally required for chromosome oscillation and checkpoint satisfaction.

    Evidence In vitro binding, co-IP, RNAi, competing peptide perturbation, and live-cell imaging of chromosome movement

    PMID:26378239

    Open questions at the time
    • Structural basis of CENP-H/CSPP1 interface not defined
    • How the interaction couples to microtubule attachment kinetics unresolved
  9. 2016 High

    Revealed a meiotic role distinct from kinetochore architecture, showing CENP-H protects cyclin B1 from APC/C(Cdh1) to drive the oocyte G2/M transition.

    Evidence Morpholino in mouse oocytes, Western blot, exogenous cyclin B1 rescue, and APC/C(Cdh1) epistasis

    PMID:27993978

    Open questions at the time
    • Direct mechanism by which CENP-H shields cyclin B1 from APC/C unknown
    • Whether this function is kinetochore-dependent unresolved
  10. 2017 Medium

    Uncovered a non-kinetochore signaling function in which CENP-H binds GOLPH3 to attenuate mTORC1/mTORC2, and placed CENP-H upstream of the intrinsic apoptotic pathway in cancer cells.

    Evidence Co-IP, GST and His pull-downs, confocal colocalization, mTOR pathway Western blot in colorectal cancer cells; siRNA knockdown with apoptosis assays and xenografts in hepatocellular carcinoma

    PMID:28498417 PMID:28819418

    Open questions at the time
    • Mechanism linking CENP-H/GOLPH3 to mTOR regulation not defined
    • Whether the signaling role is separable from the kinetochore role unclear

Open questions

Synthesis pass · forward-looking unresolved questions
  • The structural basis of CENP-H's integration into the CCAN and how its distinct kinetochore, meiotic cyclin B1, and mTOR-signaling functions are mechanistically coordinated remain unresolved.
  • No high-resolution structure of CENP-H within the inner kinetochore
  • Direct biochemical mechanism of cyclin B1 protection unknown
  • Connection between CENP-H/GOLPH3 binding and mTOR modulation undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005198 structural molecule activity 4 GO:0060090 molecular adaptor activity 2
Localization
GO:0005694 chromosome 3 GO:0000228 nuclear chromosome 2
Pathway
R-HSA-1640170 Cell Cycle 4 R-HSA-5357801 Programmed Cell Death 2 R-HSA-162582 Signal Transduction 1
Partners
CENPCCENPKCSPP1GOLPH3NDC80/HEC1TRIM36
Complex memberships
CCAN (inner kinetochore)CENP-H/CENP-K heterodimer

Evidence

Reading pass · 12 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1999 CENP-H is a constitutive kinetochore protein containing a coiled-coil structure and a nuclear localization signal, specifically and constitutively localized at kinetochores throughout the cell cycle. Protein isolation, sequence analysis, immunofluorescence with anti-CENP-H antibody The Journal of biological chemistry Medium 10488063
2001 CENP-H is required for targeting CENP-C, but not CENP-A, to the centromere in vertebrate cells; loss of CENP-H causes metaphase arrest consistent with centromere dysfunction, establishing a hierarchical centromere assembly pathway in which CENP-A localization is upstream of both CENP-C and CENP-H. Conditional loss-of-function knockout in chicken DT40 cells, immunocytochemistry The EMBO journal High 11500386
2005 The functional region of CENP-H interacts with Hec1, a member of the Nuf2 complex, both by yeast two-hybrid and coimmunoprecipitation; CENP-H and Hec1 form stable associations at centromeres during mitosis (by FRAP), suggesting the Nuf2 complex acts as a connector between inner and outer kinetochores. Yeast two-hybrid, coimmunoprecipitation, FRAP (photobleaching experiments) in chicken DT40 cells Molecular and cellular biology High 15713649
2006 RNAi knockdown of CENP-H in human HEp-2 cells to <5% of normal levels causes misaligned chromosomes and multipolar spindles, slightly reduces CENP-C levels at kinetochores, reduces CENP-E at misaligned chromosomes, but does not cause mitotic arrest and leaves hBubR1 localization normal, indicating CENP-H is required for kinetochore architecture including CENP-E recruitment. RNAi knockdown, immunofluorescence, Western blot in human HEp-2 cells Biochemical and biophysical research communications Medium 16875666
2007 CENP-H and CENP-C co-localize to discontinuous CENP-A chromatin subdomains at human neocentromeres, defining an inner kinetochore chromatin structure consistent with higher-order chromatin looping models. Chromatin immunoprecipitation on CHIP (ChIP-on-chip) using BAC and PCR-amplicon microarrays Genome biology Medium 17651496
2009 CENP-H and CENP-K form a stable ~1:1 stoichiometry subcomplex (resistant to high salt) purified by tandem affinity purification; bioinformatic analysis indicates both are enriched in coiled-coil regions, and their interacting functional regions map to their N- and C-terminals, suggesting heterodimeric coiled-coil formation within the inner kinetochore. Tandem affinity purification (TAP) from HEK293 cells expressing TAP-CENP-K, bioinformatic coiled-coil analysis Science in China. Series C, Life sciences Medium 19381461
2009 TRIM36 interacts with CENP-H (identified by yeast two-hybrid) and co-localizes with alpha-tubulin; TRIM36 has ubiquitin ligase activity and its overexpression decelerates the cell cycle, suggesting a functional link between TRIM36 and CENP-H in chromosome segregation. Yeast two-hybrid, immunofluorescence, cell cycle assay Biochemical and biophysical research communications Low 19232519
2010 Loss of cenph in zebrafish (stac mutant) causes mitotic chromosome missegregation, G2/M arrest, hyperactivation of the intrinsic apoptosis pathway (partially blocked by p53 knockdown), and embryonic lethality; heterozygosity for cenph reduces invasive tumor development, establishing CENPH as essential for mitosis and linking it to tumor suppression in vivo. Transposon insertional mutant, antisense morpholino knockdown, mRNA rescue, p53 co-knockdown epistasis, in vivo tumor incidence assay in zebrafish The Journal of biological chemistry High 20573960
2015 CSPP1 binds CENP-H both in vitro and in vivo; CSPP1 depletion impairs chromosome oscillation and spindle assembly checkpoint satisfaction similarly to CENP-H depletion; disrupting the CENP-H/CSPP1 interaction with a membrane-permeable competing peptide causes mitotic arrest and chromosome segregation defects; CSPP1 overexpression decreases kinetochore movement speed, establishing CSPP1 as a CENP-H-interacting regulator of kinetochore–microtubule dynamics. In vitro binding assay, co-immunoprecipitation, RNAi depletion, competing peptide perturbation, live-cell imaging of chromosome movement The Journal of biological chemistry High 26378239
2016 CenpH is required for meiotic G2/M transition in mouse oocytes: depletion of CenpH reduces cyclin B1 protein levels, attenuates MPF (maturation-promoting factor) activation, and severely impairs meiotic resumption; CenpH protects cyclin B1 from APC/C(Cdh1)-mediated destruction; exogenous cyclin B1 rescues the G2/M transition defect; however, CenpH depletion does not affect spindle organization or cell cycle progression after germinal vesicle breakdown. Morpholino injection in mouse oocytes, Western blot, rescue by exogenous cyclin B1 expression, epistasis with APC/CCdh1 pathway Development (Cambridge, England) High 27993978
2017 CENPH interacts physically with GOLPH3 (confirmed by co-immunoprecipitation, GST pull-down, His-tag pull-down, and confocal colocalization); through this interaction, CENPH attenuates both mTORC1 and mTORC2 signaling and reduces sensitivity to the mTOR inhibitor rapamycin in colorectal cancer cells. Co-immunoprecipitation, GST pull-down, His-tag pull-down, laser scanning confocal microscopy, Western blot for mTOR pathway components, MTT/colony formation assays Journal of Cancer Medium 28819418
2017 CENP-H knockdown in hepatocellular carcinoma Hep3B cells inhibits proliferation, induces apoptosis with activation of cleaved caspase-3, and increases Bax/Bcl-2 ratio at both mRNA and protein levels, placing CENP-H upstream of the mitochondrial (intrinsic) apoptotic pathway. siRNA knockdown, MTT assay, colony formation, transmission electron microscopy, Western blot, qRT-PCR, xenograft mouse model with IHC Oncology reports Medium 28498417

Source papers

Stage 0 corpus · 16 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2001 CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells. The EMBO journal 145 11500386
2007 Co-localization of CENP-C and CENP-H to discontinuous domains of CENP-A chromatin at human neocentromeres. Genome biology 69 17651496
1999 Characterization of a novel kinetochore protein, CENP-H. The Journal of biological chemistry 59 10488063
2009 TRIM36 interacts with the kinetochore protein CENP-H and delays cell cycle progression. Biochemical and biophysical research communications 41 19232519
2005 The functional region of CENP-H interacts with the Nuf2 complex that localizes to centromere during mitosis. Molecular and cellular biology 39 15713649
2006 Increased expression of CENP-H gene in human oral squamous cell carcinomas harboring high-proliferative activity. Oncology reports 23 17016595
2017 CENPH Inhibits Rapamycin Sensitivity by Regulating GOLPH3-dependent mTOR Signaling Pathway in Colorectal Cancer. Journal of Cancer 21 28819418
2010 Interruption of cenph causes mitotic failure and embryonic death, and its haploinsufficiency suppresses cancer in zebrafish. The Journal of biological chemistry 20 20573960
2017 CENP-H regulates the cell growth of human hepatocellular carcinoma cells through the mitochondrial apoptotic pathway. Oncology reports 19 28498417
2006 RNAi knockdown of human kinetochore protein CENP-H. Biochemical and biophysical research communications 18 16875666
2015 Mitotic Protein CSPP1 Interacts with CENP-H Protein to Coordinate Accurate Chromosome Oscillation in Mitosis. The Journal of biological chemistry 17 26378239
2016 CenpH regulates meiotic G2/M transition by modulating the APC/CCdh1-cyclin B1 pathway in oocytes. Development (Cambridge, England) 11 27993978
2012 Clinical Significance of CENP-H Expression in Uterine Cervical Cancer. Cancer biology & medicine 9 23691478
2011 The Aspergillus nidulans CENP-E kinesin motor KipA interacts with the fungal homologue of the centromere-associated protein CENP-H at the kinetochore. Molecular microbiology 9 21392133
2009 CENP-K and CENP-H may form coiled-coils in the kinetochores. Science in China. Series C, Life sciences 8 19381461
2012 Low prevalence of autoantibodies to CENP-H, -I, -K, -L, -M, -N, -T and -U in a Japanese cohort of anti-centromere positive samples. Immunopharmacology and immunotoxicology 4 23083211

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