The 19S proteasome subunit Rpt3 regulates distribution of CENP-A by associating with centromeric chromatin

Kitagawa, Teppei; Ishii, Kojiro; Takeda, Kazuhisa; Matsumoto, Tomohiro

doi:10.1038/ncomms4597

Cited by 24 publications

(34 citation statements)

References 64 publications

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“…We previously showed that mislocalization and chromosome segregation defects due to overexpression of the stable cse4 mutant, cse4 16KR (in which all 16 lysines in Cse4 are mutated to arginines), is suppressed by the constitutive expression of histone H3 (Δ16H3) [4]. Consistent with the observed increase in plasmid loss due to Cse4 mislocalization, high levels of plasmid retention (95.4%) were observed in the met30-6 strain expressing Δ16H3 at 10G (p-value = 0.08, Fig 7C), this is despite the fact that, in agreement with previous results showing an effect of altered histone stoichiometry on chromosomal stability in WT budding yeast [4] and fission yeast [69], a WT strain containing Δ16H3 showed reduced plasmid retention ( Fig 7C).…”

Section: Scf-met30 and Scf-cdc4 Prevent Mislocalization Of Cse4 To Nosupporting

confidence: 90%

Skp, Cullin, F-box (SCF)-Met30 and SCF-Cdc4-Mediated Proteolysis of CENP-A Prevents Mislocalization of CENP-A for Chromosomal Stability in Budding Yeast

Zhang

Mishra

et al. 2020

PLoS Genet

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Restricting the localization of the histone H3 variant CENP-A (Cse4 in yeast, CID in flies) to centromeres is essential for faithful chromosome segregation. Mislocalization of CENP-A leads to chromosomal instability (CIN) in yeast, fly and human cells. Overexpression and mislocalization of CENP-A has been observed in many cancers and this correlates with increased invasiveness and poor prognosis. Yet genes that regulate CENP-A levels and localization under physiological conditions have not been defined. In this study we used a genome-wide genetic screen to identify essential genes required for Cse4 homeostasis to prevent its mislocalization for chromosomal stability. We show that two Skp, Cullin, Fbox (SCF) ubiquitin ligases with the evolutionarily conserved F-box proteins Met30 and Cdc4 interact and cooperatively regulate proteolysis of endogenous Cse4 and prevent its mislocalization for faithful chromosome segregation under physiological conditions. The interaction of Met30 with Cdc4 is independent of the D domain, which is essential for their homodimerization and ubiquitination of other substrates. The requirement for both Cdc4 and Met30 for ubiquitination is specifc for Cse4; and a common substrate for Cdc4 and Met30 has not previously been described. Met30 is necessary for the interaction between Cdc4 and Cse4, and defects in this interaction lead to stabilization and mislocalization of Cse4, which in turn contributes to CIN. We provide the first direct link between Cse4 mislocalization to defects in kinetochore structure and show that SCF-mediated proteolysis of

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Section: Scf-met30 and Scf-cdc4 Prevent Mislocalization Of Cse4 To Nosupporting

confidence: 90%

Skp, Cullin, F-box (SCF)-Met30 and SCF-Cdc4-Mediated Proteolysis of CENP-A Prevents Mislocalization of CENP-A for Chromosomal Stability in Budding Yeast

Zhang

Mishra

et al. 2020

PLoS Genet

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“…To test if our screen identified the same set of extragenic suppressor genes as Li et al, we determined the whole-genome sequences of 3 BOE strains for mrpl8 (mitochondrial ribosome protein), which was a C-BOE hit in Li et al (2019). Interestingly, we identified mutations in atp1 (F1-F0 ATP synthase alpha subunit (Falson et al ., 1991)) and rpt3 (19S proteasome base subcomplex ATPase subunit (Kitagawa et al ., 2014)), which are very similar to what were found in Li et al (2019) ( atp3; F1-F0 ATP synthase gamma subunit: mts4; 19S proteasome regulatory subunit (Wilkinson et al ., 1997)). In addition, our screen uniquely identified hul5 (HECT-type ubiquitin-protein ligase E3 (Fang et al ., 2011)), which might function upstream of the proteasome.…”

Section: Resultsmentioning

confidence: 99%

Identification of 15 new bypassable essential genes of fission yeast

Takeda

Saitoh

Ohkura

et al. 2019

Preprint

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15Every organism has a different set of genes essential for its viability. This indicates that an 16 organism can become tolerant to the loss of an essential gene under certain circumstances during 17 evolution, via the manifestation of 'masked' alternative mechanisms. In our quest to 18 systematically uncover masked mechanisms in eukaryotic cells, we developed an extragenic 19 suppressor screening method using haploid spores deleted of an essential gene in the fission yeast 20 Schizosaccharomyces pombe. We screened for the 'bypass' suppressors of lethality of 92 21 randomly selected genes that are essential for viability in standard laboratory culture conditions. 22Remarkably, extragenic mutations bypassed the essentiality of as many as 20 genes (22%), 15 of 23 which have not been previously reported. Half of the bypass-suppressible genes were involved in 24 mitochondria function; we also identified multiple genes regulating RNA processing. 18 25 suppressible genes were conserved in the budding yeast Saccharomyces cerevisiae, but 13 of them 26 were non-essential in that species. These trends are consistent with a recent independent 27 bypass-of-essentiality (BOE) screening of 142 fission yeast genes conducted with more elaborate 28 methodology (Li et al., 2019). Thus, our study reinforces the emerging view that BOE is not a rare 29 event and that each organism may be endowed with secondary or backup mechanisms that can 30 substitute for primary mechanisms in various biological processes. Furthermore, the robustness of 31 our simple spore-based methodology paves the way for genome-scale BOE screening. 32 33

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“…Several studies have shown the presence of the proteasome in eukaryotic nuclei [40–44] and its recruitment to chromatin including centromeres [45], telomeres [46] and sites of cryptic transcriptional initiation [38]. To investigate whether the proteasome might participate in transcriptional silencing of heterochromatin, proteasome binding at pericentromeric and several other endogenous repeats was analysed using ChIP-seq data previously obtained in mouse 3T3-L1 cells [47].…”

Section: Resultsmentioning

confidence: 99%

Transcriptional Activation of Pericentromeric Satellite Repeats and Disruption of Centromeric Clustering upon Proteasome Inhibition

et al. 2016

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Heterochromatinisation of pericentromeres, which in mice consist of arrays of major satellite repeats, are important for centromere formation and maintenance of genome stability. The dysregulation of this process has been linked to genomic stress and various cancers. Here we show in mice that the proteasome binds to major satellite repeats and proteasome inhibition by MG132 results in their transcriptional de-repression; this de-repression is independent of cell-cycle perturbation. The transcriptional activation of major satellite repeats upon proteasome inhibition is accompanied by delocalisation of heterochromatin protein 1 alpha (HP1α) from chromocentres, without detectable change in the levels of histone H3K9me3, H3K4me3, H3K36me3 and H3 acetylation on the major satellite repeats. Moreover, inhibition of the proteasome was found to increase the number of chromocentres per cell, reflecting destabilisation of the chromocentre structures. Our findings suggest that the proteasome plays a role in maintaining heterochromatin integrity of pericentromeres.

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The 19S proteasome subunit Rpt3 regulates distribution of CENP-A by associating with centromeric chromatin

Cited by 24 publications

References 64 publications

Skp, Cullin, F-box (SCF)-Met30 and SCF-Cdc4-Mediated Proteolysis of CENP-A Prevents Mislocalization of CENP-A for Chromosomal Stability in Budding Yeast

Skp, Cullin, F-box (SCF)-Met30 and SCF-Cdc4-Mediated Proteolysis of CENP-A Prevents Mislocalization of CENP-A for Chromosomal Stability in Budding Yeast

Identification of 15 new bypassable essential genes of fission yeast

Transcriptional Activation of Pericentromeric Satellite Repeats and Disruption of Centromeric Clustering upon Proteasome Inhibition

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