N-terminal sumoylation of centromeric histone H3 variant Cse4 regulates its proteolysis to prevent mislocalization to non-centromeric chromatin

Ohkuni, Kentaro; Levy-Myers, Reuben; Warren, Jack; Au, Wei-Chun; Takahashi, Yoshimitsu; Baker, Richard E.; Basrai, Munira A.

doi:10.1101/176206

Cited by 11 publications

(13 citation statements)

References 37 publications

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“…Wild type, hhf1Δ, and hhf2Δ GALCSE4 strains were assayed for Cse4 sumoylation. Consistent with previous results (OHKUNI et al 2016;OHKUNI et al 2018;OHKUNI et al 2020), we detected sumoylated Cse4 as a pattern of three high molecular weight bands in wild type cells overexpressing wild type Cse4 but not in wild type cells expressing vector alone or overexpressing cse4 16KR ( Figure 5A). Deletion of either histone H4 allele resulted in reduced levels of sumoylated Cse4 (Figures 5A and 5B; p-value WT vs hhf1Δ = 0.0006, p-value WT vs hhf2Δ = 0.0007).…”

Section: Reduced Dosage Of H4 Is Associated With Defects In Sumoylatisupporting

confidence: 92%

Reduced Gene Dosage of Histone H4 Prevents CENP-A Mislocalization inSaccharomyces cerevisiae

Eisenstatt

Ohkuni

et al. 2020

Preprint

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Mislocalization of the centromeric histone H3 variant (Cse4 in budding yeast, CID in flies, CENP-A in humans) contributes to chromosomal instability (CIN) in yeast, fly, and human cells. Overexpression and mislocalization of CENP-A has been observed in several cancers. However, the mechanisms that contribute to the mislocalization of CENP-A are not fully understood. In this study, we used budding yeast to identify genes that facilitate the mislocalization of Cse4 to non-centromeric regions. Previous studies have shown that E3 ligases (Psh1, Slx5, Cdc4) and factors such as Doa1, Hir2, and Cdc7 regulate proteolysis of Cse4 and prevent its mislocalization. Overexpressed Cse4 (GALCSE4) is highly stable and causes synthetic dosage lethality (SDL) in psh1Δ, slx5Δ, doa1Δ, hir2Δ, cdc4-1, and cdc7-4 strains. We used a genome-wide screen to identify suppressors of the psh1Δ GALCSE4 SDL. Deletions of histone H4 alleles (HHF1 or HHF2) were among the top suppressors of psh1Δ GALCSE4 SDL. Here we show that reduced gene dosage of H4 prevents mislocalization of Cse4 and promotes faster degradation of Cse4 in a psh1Δ GALCSE4 strain. Deletion of either HHF1 or HHF2 also suppresses the GALCSE4 SDL in slx5Δ, doa1Δ, hir2Δ, cdc4-1, and cdc7-4 strains. Suppression of psh1Δ GALCSE4 SDL by hhf1-20, which is defective for interaction with Cse4, suggests that defects in the Cse4-H4 interaction prevents mislocalization of Cse4. In summary, our genome-wide screen identified genes that contribute to Cse4 mislocalization and we show how reduced dosage of histone H4-encoding genes prevents mislocalization of Cse4 into non-centromeric regions.

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Section: Reduced Dosage Of H4 Is Associated With Defects In Sumoylatisupporting

confidence: 92%

Reduced Gene Dosage of Histone H4 Prevents CENP-A Mislocalization inSaccharomyces cerevisiae

Eisenstatt

Ohkuni

et al. 2020

Preprint

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“…Indeed, the misregulation of mouse minor satellite RNAs leads to impaired centromere function and defective chromosome segregation [ 35 ]. In addition, work in budding yeast has focused on how CENP-A Cse4 loading is limited to the centromere [ 143 , 144 , 145 , 146 , 147 ]. Whether the removal of ectopic CENP-A in non-Ascomycota species happens by the same mechanism with the same efficiency remains to be determined.…”

Section: Centromeric Transcription In Diseasementioning

confidence: 99%

“…One would expect that mechanisms have evolved to reduce this risk to a minimum without impacting the native centromere functions. In the case of the point centromere of budding yeast, which is thought to contain only of a single CENP-A Cse4 nucleosome [ 158 ], SUMOylation of N-terminal tail of CENP-A Cse4 is a driving force in preventing CENP-A Cse4 mislocalization [ 143 , 144 , 145 , 146 , 147 ]. In plants, active removal of CENP-A Cse4 has also been observed [ 159 ].…”

Section: Centromeric Transcription In Diseasementioning

confidence: 99%

Centromeric Transcription: A Conserved Swiss-Army Knife

Arunkumar

Melters

2020

Genes

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In most species, the centromere is comprised of repetitive DNA sequences, which rapidly evolve. Paradoxically, centromeres fulfill an essential function during mitosis, as they are the chromosomal sites wherein, through the kinetochore, the mitotic spindles bind. It is now generally accepted that centromeres are transcribed, and that such transcription is associated with a broad range of functions. More than a decade of work on this topic has shown that centromeric transcripts are found across the eukaryotic tree and associate with heterochromatin formation, chromatin structure, kinetochore structure, centromeric protein loading, and inner centromere signaling. In this review, we discuss the conservation of small and long non-coding centromeric RNAs, their associations with various centromeric functions, and their potential roles in disease.

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“…As a deSUMOylase, SENP6 could prevent CENP-I, and other centromere/kinetochore protein, degradation by removing SUMO chains. Indeed, SUMO-mediated polyubiquitination and subsequent degradation of budding yeast CENP-A Cse4 has been reported (Ohkuni et al, 2018(Ohkuni et al, , 2016. Therefore, we measured the protein stability of CENP-A and -C after depletion of SENP6.…”

Section: Senp6 Is Involved In Maintaining the Entire Centromere Complmentioning

confidence: 97%

Genetic screening identifies a SUMO protease dynamically maintaining centromeric chromatin and the associated centromere complex

Mitra

Bodor

David

et al. 2019

Preprint

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Centromeres are defined by a unique self-propagating chromatin structure featuring nucleosomes containing the histone H3 variant CENP-A. CENP-A turns over slower than general chromatin and a key question is whether this unusual stability is intrinsic to CENP-A nucleosomes or rather imposed by external factors. We designed a specific genetic screen to identify proteins involved in CENP-A stability based on SNAP-tag pulse chase labeling. Using a double pulse-labeling approach we simultaneously assay for factors with selective roles in CENP-A chromatin assembly. We discover a series of new proteins involved in CENP-A propagation, including proteins with known roles in DNA replication, repair and chromatin modification and transcription, revealing that a broad set of chromatin regulators impacts in CENP-A transmission through the cell cycle.The key factor we find to strongly affect CENP-A stability is SENP6. This SUMO-protease controls not only the levels of chromatin bound CENP-A but is required for the maintenance of virtually the entire centromere and kinetochore, with the exception of CENP-B. Acute depletion of SENP6 protein reveals its requirement for maintaining centromeric CENP-A levels throughout the cell cycle, suggesting that a dynamic SUMO cycle underlies a continuous surveillance of the centromere complex.

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N-terminal sumoylation of centromeric histone H3 variant Cse4 regulates its proteolysis to prevent mislocalization to non-centromeric chromatin

Cited by 11 publications

References 37 publications

Reduced Gene Dosage of Histone H4 Prevents CENP-A Mislocalization inSaccharomyces cerevisiae

Reduced Gene Dosage of Histone H4 Prevents CENP-A Mislocalization inSaccharomyces cerevisiae

Centromeric Transcription: A Conserved Swiss-Army Knife

Genetic screening identifies a SUMO protease dynamically maintaining centromeric chromatin and the associated centromere complex

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