Abstract:
:In Escherichia coli, ClpYQ (HslUV) is a two-component ATP-dependent protease composed of ClpY (HslU), an ATPase with unfolding activity, and ClpQ (HslV), a peptidase. In the ClpYQ proteolytic complex, the hexameric rings of ClpY (HslU) are responsible for protein recognition, unfolding, and translocation into the proteolytic inner chamber of the dodecameric ClpQ (HslV). Each of the three domains, N, I, and C, in ClpY has its own distinct activity. The double loops (amino acids [aa] 137 to 150 and 175 to 209) in domain I of ClpY are necessary for initial recognition/tethering of natural substrates such as SulA, a cell division inhibitor protein. The highly conserved sequence GYVG (aa 90 to 93) pore I site, along with the GESSG pore II site (aa 265 to 269), contribute to the central pore of ClpY in domain N. These two central loops of ClpY are in the center of its hexameric ring in which the energy of ATP hydrolysis allows substrate translocation and then degradation by ClpQ. However, no data have been obtained to determine the effect of the central loops on substrate binding or as part of the processivity of the ClpYQ complex. Thus, we probed the features of ClpY important for substrate engagement and protease processivity via random PCR or site-specific mutagenesis. In yeast two-hybrid analysis and pulldown assays, using isolated ClpY mutants and the pore I or pore II site of ClpY, each was examined for its influence on the adjoining structural regions of the substrates. The pore I site is essential for the translocation of the engaged substrates. Our in vivo study of the ClpY mutants also revealed that an ATP-binding site in domain N, separate from its role in polypeptide (ClpY) oligomerization, is required for complex formation with ClpQ. Additionally, we found that the tyrosine residue at position 408 in ClpY is critical for stabilization of hexamer formation between subunits. Therefore, our studies suggest that stepwise activities of the ClpYQ protease are necessary to facilitate the processive degradation of its natural substrates.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Hsieh FC,Chen CT,Weng YT,Peng SS,Chen YC,Huang LY,Hu HT,Wu YL,Lin NC,Wu WFdoi
10.1128/JB.05128-11subject
Has Abstractpub_date
2011-10-01 00:00:00pages
5465-76issue
19eissn
0021-9193issn
1098-5530pii
JB.05128-11journal_volume
193pub_type
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journal_title:Journal of bacteriology
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doi:10.1128/JB.117.3.1178-1183.1974
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doi:10.1128/jb.172.1.224-235.1990
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journal_title:Journal of bacteriology
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doi:10.1128/JB.181.16.4896-4904.1999
更新日期:1999-08-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/jb.173.18.5604-5611.1991
更新日期:1991-09-01 00:00:00
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journal_title:Journal of bacteriology
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00188-18
更新日期:2018-08-10 00:00:00
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pub_type: 杂志文章
doi:10.1128/JB.143.2.1019-1024.1980
更新日期:1980-08-01 00:00:00
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pub_type: 杂志文章
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.87.2.446-453.1964
更新日期:1964-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.171.9.4640-4647.1989
更新日期:1989-09-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/jb.175.23.7617-7623.1993
更新日期:1993-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2012-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2003-07-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1972-10-01 00:00:00
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更新日期:1994-10-01 00:00:00
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pub_type: 杂志文章
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更新日期:1987-09-01 00:00:00
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journal_title:Journal of bacteriology
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doi:10.1128/JB.130.1.107-113.1977
更新日期:1977-04-01 00:00:00
abstract::A third nuclear protein-coding gene termed PET122 has been shown to be required for a post-transcriptional step in expression of the mitochondrial COX3 gene is Saccharomyces cerevisiae. pet122 mutants fail to produce cytochrome c oxidase subunit III, which is the polypeptide product of the COX3 gene, but produce norma...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.3.1399-1402.1988
更新日期:1988-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.169.1.371-379.1987
更新日期:1987-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01194-07
更新日期:2008-06-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.115.3.746-751.1973
更新日期:1973-09-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:2000-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.181.15.4719-4723.1999
更新日期:1999-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.169.8.3414-3421.1987
更新日期:1987-08-01 00:00:00
abstract::When soluble extracts of the extreme acidothermophilic archaeon Sulfolobus solfataricus were incubated with [gamma-(32)P]ATP, several proteins were radiolabeled. One of the more prominent of these, which migrated with a mass of approximately 46 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE...
journal_title:Journal of bacteriology
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更新日期:2003-04-01 00:00:00
abstract::In Acinetobacter calcoaceticus the seven genes coding for the enzymes responsible for tryptophan synthesis map at three chromosomal locations. Two three-gene clusters, one (trpGDC) specifying the small subunit of anthranilate synthase, phosphoribosyl transferase, and indoleglycerol phosphate synthase and the other (tr...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.127.1.367-379.1976
更新日期:1976-07-01 00:00:00