Quiescent Cells Actively Replenish CENP-A Nucleosomes to Maintain Centromere Identity and Proliferative Potential

Swartz, S. Zachary; McKay, Liliana S.; Su, Kuan-Chung; Bury, Leah; Padeganeh, Abbas; Maddox, Paul S.; Knouse, Kristin A.; Cheeseman, Iain M.

doi:10.1016/j.devcel.2019.07.016

Cited by 72 publications

(50 citation statements)

References 45 publications

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“…We find that the predominant factor controlling alpha-satellite transcription is the presence of centromere-nucleolar contacts, providing a mechanism to modulate centromere transcription and chromatin dynamics across diverse cell states and conditions. Together, our results support a role for ongoing transcription in promoting the dynamics of centromeric chromatin (Swartz et al, 2019).…”

Section: Introductionsupporting

confidence: 71%

“…The nature of the behavior that we observed for alphasatellite smFISH foci, including the lack of localization to centromeres or mitotic structures, is inconsistent with a direct, physical role for these transcripts in cell division processes. Instead, this suggests a role for the transcription process itself, such as a role for ongoing transcription to promote the dynamics and rejuvenation of centromeric chromatin, as proposed by our recent work in non-dividing cells (Swartz et al, 2019).…”

Section: Nucleolar Associations Act To Repress Alpha-satellite Transcmentioning

confidence: 85%

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Alpha-satellite RNA transcripts are repressed by centromere-nucleolus associations

Bury

Moodie

McKay

et al. 2020

Preprint

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Centromeres play a fundamental role in chromosome segregation. Although originally thought to be silent chromosomal regions, centromeres are actively transcribed.However, the behavior and contributions of centromere-derived RNAs have remained unclear. Here, we used single-molecule fluorescence in-situ hybridization (smFISH) to detect alpha-satellite RNA transcripts in intact human cells. We find that alpha-satellite RNA smFISH foci fluctuate in their levels over the cell cycle and do not remain associated with centromeres, displaying localization consistent with other long noncoding RNAs. Our results demonstrate that alpha-satellite expression occurs through RNA Polymerase II-dependent transcription, but does not require centromere proteins and other cell division components. Instead, our work implicates centromere-nucleolar associations as the major factor regulating alpha-satellite expression. The fraction of nucleolar-localized centromeres inversely correlates with alpha-satellite transcripts levels, explaining variations in alpha-satellite RNA between cell lines. In addition, alphasatellite transcript levels increase substantially when the nucleolus is disrupted.Together, our results are inconsistent with a direct, physical role for alpha-satellite transcripts in cell division processes, and instead support a role for ongoing transcription in promoting centromere chromatin dynamics. The control of alpha-satellite transcription by centromere-nucleolar contacts provides a mechanism to modulate centromere transcription and chromatin dynamics across diverse cell states and conditions. Alexandrov, I., A. Kazakov, I. Tumeneva, V. Shepelev, and Y. Yurov. 2001. Alphasatellite DNA of primates: old and new families. Chromosoma. 110:253-266. . 2012. Epigenetic engineering: histone H3K9 acetylation is compatible with kinetochore structure and function. J Cell Sci. 125:411-421. . 2011. Epigenetic engineering shows H3K4me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore. Embo J. 30:328-340. Biscotti, M.A., A. Canapa, M. Forconi, E. Olmo, and M. Barucca. 2015. Transcription of tandemly repetitive DNA: functional roles.

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Section: Introductionsupporting

confidence: 71%

Section: Nucleolar Associations Act To Repress Alpha-satellite Transcmentioning

confidence: 85%

Alpha-satellite RNA transcripts are repressed by centromere-nucleolus associations

Bury

Moodie

McKay

et al. 2020

Preprint

Self Cite

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“…This frequent mis-segregation suggested a more severe defect, perhaps due to active stripping of previously loaded CENP-A from the centromeres of bridge chromosomes. To address this possibility, we pulse-labeled cells expressing Halo-tagged CENP-A from its endogenous locus (55) prior to the induction of chromosome bridges, enabling preferential visualization of the preexisting population of CENP-A that was loaded prior to bridge formation. After labeling and bridge induction (TRF2-DN), cells were given sufficient time to divide twice—first to form bridges, and again to allow bridges to be converted to micronuclei (Fig.…”

Section: Resultsmentioning

confidence: 99%

Mechanisms Generating Cancer Genome Complexity From A Single Cell Division Error

Umbreit

Chen

Lynch

et al. 2019

Preprint

154

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ABSTRACTThe chromosome breakage-fusion-bridge (BFB) cycle is a mutational process that produces gene amplification and genome instability. Signatures of BFB cycles can be observed in cancer genomes with chromothripsis, another catastrophic mutational process. Here, we explain this association by identifying a mutational cascade downstream of chromosome bridge formation that generates increasing amounts of chromothripsis. We uncover a new role for actomyosin forces in bridge breakage and mutagenesis. Chromothripsis then accumulates starting with aberrant interphase replication of bridge DNA, followed by an unexpected burst of mitotic DNA replication, generating extensive DNA damage. Bridge formation also disrupts the centromeric epigenetic mark, leading to micronucleus formation that itself promotes chromothripsis. We show that this mutational cascade generates the continuing evolution and sub-clonal heterogeneity characteristic of many human cancers.

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“…These results suggest that both Cid1 and Cid5 are present at centromeres early in oogenesis but only Cid1 remains on centromeres by meiosis I metaphase arrest. Given that turnover of CenH3-containing nucleosomes in MI-arrested oocytes appears to be quite gradual (~2% of centromeric CenH3 is exchanged per day in MI-arrested starfish oocytes (Swartz, Mckay et al 2018)), we hypothesize that Cid5 protein is actively removed from the oocyte centromeres and at the onset of meiosis I metaphase arrest. Since Cid1 is always present throughout oogenesis, it is unclear whether Cid5 performs any function, centromeric or otherwise, in the female germline.…”

Section: Differential Localization Of Cid1 and Cid5 In D Virilis Ovamentioning

confidence: 96%

“…Oocyte centromere function does not seem to depend on the loading of newly transcribed CenH3 as conditional knockouts of CenH3 in meiotic prophase I are fully fertile in Mus musculus (Smoak, Stein et al 2016). However, recent work demonstrated that CenH3 in MI arrested starfish oocytes undergoes gradual turnover, presumably to replace CenH3 containing nucleosomes that are disturbed by transcriptional machinery, allowing oocytes to maintain centromere competence over long periods of time (Swartz, Mckay et al 2018). This means that CenH3 molecules are capable of stably persisting in oocytes for long periods of time and that there are mechanisms in place to maintain centromere function in non-dividing cells.…”

Section: Introductionmentioning

confidence: 99%

Gametic specialization of centromeric histone paralogs inDrosophila virilis

Kursel

Malik

2019

Preprint

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In most eukaryotes, centromeric histone (CenH3) proteins mediate the highly conserved process of chromosome segregation as the foundational kinetochore assembly factor. However, in multicellular organisms, CenH3 proteins have to perform their essential functions in different chromatin environments. CenH3 proteins not only mediate mitosis and meiosis but also ensure epigenetic inheritance of centromere identity on sperm chromatin, which is highly compact and almost completely stripped of histones during spermiogenesis. We hypothesized that such disparate chromatin environments might impose different functional constraints on CenH3. If so, gene duplications could ameliorate the difficulty of encoding divergent and even potentially incompatible centromeric functions in the same gene. Here, we analyzed the cytological localization of two recently identified CenH3 paralogs, Cid1 and Cid5, in D. virilis using specific antibodies and epitope-tagged transgenic strains. We find that only ancestral Cid1 is present in somatic cells, whereas both Cid1 and Cid5 are expressed in testes and ovaries. However, Cid1 and Cid5 are alternately retained in male and female gametes; Cid1 is lost in male meiosis but retained throughout oogenesis, whereas Cid5 is lost during female meiosis but retained in mature sperm. Following fertilization, maternally deposited Cid1 rapidly replaces paternal Cid5 during the protamine-to-histone transition. Our studies reveal mutually exclusive gametic specialization of two divergent CenH3 paralogs. We suggest that centromeric histone duplication and divergence may allow essential genes involved in chromosome segregation to specialize and thereby resolve an intralocus conflict between maternal and paternal centromeric histone requirements in many animal species.

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Quiescent Cells Actively Replenish CENP-A Nucleosomes to Maintain Centromere Identity and Proliferative Potential

Cited by 72 publications

References 45 publications

Alpha-satellite RNA transcripts are repressed by centromere-nucleolus associations

Alpha-satellite RNA transcripts are repressed by centromere-nucleolus associations

Mechanisms Generating Cancer Genome Complexity From A Single Cell Division Error

Gametic specialization of centromeric histone paralogs inDrosophila virilis

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