Phosphorylation by Casein Kinase 2 Facilitates Psh1 Protein-assisted Degradation of Cse4 Protein

Hewawasam, Geetha S.; Mattingly, Mark; Venkatesh, Swaminathan; Zhang, Ying; Florens, Laurence; Workman, Jerry L.; Gerton, Jennifer L.

doi:10.1074/jbc.m114.580589

Cited by 24 publications

(24 citation statements)

References 38 publications

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“…To determine the functional relationship between PSH1 and RCY1, we assessed the degradation of Cse4 in cells lacking either or both RCY1 and PSH1. Consistent with earlier studies, deletion of PSH1 slightly stabilized Cse4 (11,12,14,15). RCY1 deletion appeared to cause stronger Cse4 stabilization (Fig.…”

Section: Resultssupporting

confidence: 80%

“…Consistent with previous studies, Cse4 degradation is compromised in yeast cells missing PSH1 or DOA1 (11,12,14,15). Four isolated mutants (i.e.…”

Section: Resultssupporting

confidence: 73%

“…As Cse4 is still degraded in yeast cells lacking Psh1 E3 (11)(12)(13)(14)(15), other ubiquitin ligases likely are involved in Cse4 turnover. To this end, we employed an array of yeast mutant cells lacking known or putative ubiquitylation/degradation enzymes (22) (supplemental Table S1).…”

Section: Resultsmentioning

confidence: 99%

“…The mechanism underlying Psh1-mediated Cse4 turnover remains to be elucidated. Because Cse4 degradation is partially impaired in psh1⌬ cells, other E3s likely also promote Cse4 turnover (11)(12)(13)(14)(15).…”

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confidence: 99%

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The F-box Protein Rcy1 Is Involved in the Degradation of Histone H3 Variant Cse4 and Genome Maintenance

Cheng

Bao

Rao

2016

Journal of Biological Chemistry

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Cse4, a histone H3-like centromeric protein, plays critical functions in chromosome segregation. Cse4 level is tightly regulated, but the underlying mechanism remains poorly understood. We employed a toxicity-based screen to look for the degradation components involved in Cse4 regulation. Here, we show that the F-box containing protein Rcy1 is required for efficient Cse4 turnover as Cse4 degradation is compromised in yeast cells lacking RCY1. Excessive Cse4 accumulation in rcy1⌬ cells leads to growth retardation. Furthermore, the deletion of RCY1 is tied to enhanced chromosome instability and temperature-sensitive cell growth. Our results reveal the involvement of Rcy1 in chromosome regulation and another regulatory pathway controlling the Cse4 level and activity.Chromosome segregation is key to equally divide the duplicated DNA to daughter cells. Following DNA replication, sister chromatids are joined at a highly condensed and constricted chromosome region called centromere (1,2). A distinct feature of the specialized centromeric chromatin is the incorporation of an essential histone H3 variant CENP-A (called Cse4 in yeast Saccharomyces cerevisiae), instead of histone H3 in nucleosomes (1,3,4).As altered localization and expression of CENP-A and its homologues have been linked to genome instability and cancers (1,5,6), CENP-A activity must be tightly regulated. Human CENP-A and yeast Cse4 have been found to be subject to proteasome-mediated degradation (1,7,8). Proteasome substrates are selected by a ubiquitin ligase E3, which collaborates with a ubiquitin-activating enzyme (E1) and a ubiquitin-conjugating enzyme (E2) to covalently attach ubiquitin molecules onto substrates (9, 10). Ubiquitin-marked substrates are then delivered to and degraded by the proteasome. The key to understanding the role of proteolysis in CENP-A regulation is to identify E3 enzymes involved.Two groups previously isolated Psh1 as a yeast Cse4-binding protein via the affinity purification-mass spectrometry coupled approach and further demonstrated that Psh1 acts as an E3 ligase for Cse4 ubiquitylation and degradation (11,12). The mechanism underlying Psh1-mediated Cse4 turnover remains to be elucidated. Because Cse4 degradation is partially impaired in psh1⌬ cells, other E3s likely also promote Cse4 turnover (11-15).We have previously developed a synthetic dosage lethality screen to uncover physiological targets of several proteolytic pathways (16 -18). The synthetic lethality screen can be easily adapted to isolate relevant components involved in the degradation of a given substrate. Because Cse4 overexpression leads to toxicity in psh1⌬ cells (11, 12), we looked for the degradation mutants that grow slower upon Cse4 overexpression. Yeast cells lacking RCY1 are found to be sensitive to Cse4 expression and exhibit compromised Cse4 degradation. RCY1 encodes a F-box containing protein, which is a substrate recognition component of the Skp1-Cdc53-F box (SCF) 2 E3 complex (19). Rcy1 is one of the eight F-box proteins in yeast, but little is ...

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Section: Resultssupporting

confidence: 80%

“…Consistent with previous studies, Cse4 degradation is compromised in yeast cells missing PSH1 or DOA1 (11,12,14,15). Four isolated mutants (i.e.…”

Section: Resultssupporting

confidence: 73%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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The F-box Protein Rcy1 Is Involved in the Degradation of Histone H3 Variant Cse4 and Genome Maintenance

Cheng

Bao

Rao

2016

Journal of Biological Chemistry

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“…Despite Psh1 localization to centromeres, centromeric Cse4 is precluded from ubiquitylation through its interaction with the Scm3 chaperone (Hewawasam et al 2010). Psh1 activity towards Cse4 is facilitated by casein kinase 2-mediated phosphorylation and its association with the chromatin modifying FACT (facilitates chromatin transcription/transactions) complex (Deyter and Biggins 2014; Hewawasam et al 2014), providing additional levels of regulation.…”

Section: Diversity Of Cenp-a Modifications Across Speciesmentioning

confidence: 99%

Posttranslational modifications of CENP-A: marks of distinction

Srivastava

Foltz

2018

Chromosoma

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Centromeres are specialized chromosome domain that serve as the site for kinetochore assembly and microtubule attachment during cell division, to ensure proper segregation of chromosomes. In higher eukaryotes, the identity of active centromeres is marked by the presence of CENP-A (centromeric protein-A), a histone H3 variant. CENP-A forms a centromere-specific nucleosome that acts as a foundation for centromere assembly and function. The posttranslational modification (PTM) of histone proteins is a major mechanism regulating the function of chromatin. While a few CENP-A site-specific modifications are shared with histone H3, the majority are specific to CENP-A-containing nucleosomes, indicating that modification of these residues contribute to centromere-specific function. CENP-A undergoes posttranslational modifications including phosphorylation, acetylation, methylation, and ubiquitylation. Work from many laboratories have uncovered the importance of these CENP-A modifications in its deposition at centromeres, protein stability, and recruitment of the CCAN (constitutive centromere-associated network). Here, we discuss the PTMs of CENP-A and their biological relevance.

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