Abstract:
:We characterized a temperature-sensitive mutant of Saccharomyces cerevisiae in which a mini-chromosome was unstable at a high temperature and cloned a new gene which encodes a basic and hydrophilic protein (110 kDa). The disruption of this gene caused the same temperature-sensitive growth as the original mutation. By using the two-hybrid system, we further isolated RSP5 (reverses Spt- phenotype), which encodes a hect (homologous to E6-AP C terminus) domain, as a gene encoding a ubiquitin ligase. Thus, we named our gene BUL1 (for a protein that binds to the ubiquitin ligase). BUL1 seems to be involved in the ubiquitination pathway, since a high dose of UBI1, encoding a ubiquitin, partially suppressed the temperature sensitivity of the bul1 disruptant as well as that of a rsp5 mutant. Coexpression of RSP5 and BUL1 on a multicopy plasmid was toxic for mitotic growth of the wild-type cells. Pulse-chase experiments revealed that Bul1 in the wild-type cells remained stable, while the bands of Bul1 in the rsp5 cells were hardly detected. Since the steady-state levels of the protein were the same in the two strains as determined by immunoblotting analysis, Bul1 might be easily degraded during immunoprecipitation in the absence of intact Rsp5. Furthermore, both Bul1 and Rsp5 appeared to be associated with large complexes which were separated through a sucrose gradient centrifugation, and Rsp5 was coimmunoprecipitated with Bul1. We discuss the possibility that Bul1 functions together with Rsp5 in protein ubiquitination.
journal_name
Mol Cell Bioljournal_title
Molecular and cellular biologyauthors
Yashiroda H,Oguchi T,Yasuda Y,Toh-E A,Kikuchi Ydoi
10.1128/mcb.16.7.3255subject
Has Abstractpub_date
1996-07-01 00:00:00pages
3255-63issue
7eissn
0270-7306issn
1098-5549journal_volume
16pub_type
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journal_title:Molecular and cellular biology
pub_type: 杂志文章
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pub_type: 杂志文章
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journal_title:Molecular and cellular biology
pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1996-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:2012-12-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1986-12-01 00:00:00
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journal_title:Molecular and cellular biology
pub_type: 杂志文章
doi:10.1128/mcb.16.8.4024
更新日期:1996-08-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:2000-10-01 00:00:00
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journal_title:Molecular and cellular biology
pub_type: 杂志文章
doi:10.1128/MCB.26.5.1806-1816.2006
更新日期:2006-03-01 00:00:00
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journal_title:Molecular and cellular biology
pub_type: 杂志文章
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更新日期:2011-04-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:1984-12-01 00:00:00
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pub_type: 杂志文章
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更新日期:1994-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:2012-07-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/mcb.6.2.539
更新日期:1986-02-01 00:00:00
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pub_type: 杂志文章
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更新日期:2005-07-01 00:00:00
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pub_type: 杂志文章
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更新日期:1998-05-01 00:00:00