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Cloning and functional analysis of the ubiquitin-specific protease gene UBP1 of Saccharomyces cerevisiae

Tobias, John W. and Varshavsky, Alexander (1991) Cloning and functional analysis of the ubiquitin-specific protease gene UBP1 of Saccharomyces cerevisiae. Journal of Biological Chemistry, 266 (18). pp. 12021-12028. ISSN 0021-9258. doi:10.1016/s0021-9258(18)99059-9.

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In eukaryotes, both natural and engineered fusions of ubiquitin to itself or other proteins are cleaved by processing proteases after the last (Gly76) residue of ubiquitin. Using the method of sib selection, and taking advantage of the fact that bacteria such as Escherichia coli lack ubiquitin-specific enzymes, we have cloned a gene, named UBP1, of the yeast Saccharomyces cerevisiae that encodes a ubiquitin-specific processing protease. With the exception of polyubiquitin, the UBP1 protease cleaves at the carboxyl terminus of the ubiquitin moiety in natural and engineered fusions irrespective of their size or the presence of an amino-terminal ubiquitin extension. These properties of UBP1 distinguish it from the previously cloned yeast protease YUH1, which deubiquitinates relatively short ubiquitin fusions but is virtually inactive with longer fusions such as ubiquitin-beta-galactosidase. The amino acid sequence of the 809-residue UBP1 lacks significant similarities to other known proteins, including the 236-residue YUH1 protease. Null ubp1 mutants are viable, and retain the ability to deubiquitinate ubiquitin-beta-galactosidase, indicating that the family of ubiquitin-specific proteases in yeast is not limited to UBP1 and YUH1.

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Varshavsky, Alexander0000-0002-4011-258X
Additional Information:© 1991 ASBMB. User License: Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0). (Received for publication, February 5, 1991). This work was supported by Grants AG08991 and DK39520 (to A. V.) from the National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. We thank Vincent Chau for the anti-Ub antibody and Rohan Baker for the pRBUl plasmid. We also thank Erica Johnson, Tom Shrader, and especially Rohan Baker for comments on the manuscript and Barbara Doran for secretarial assistance. The nucleotide sequence(s) reported in this paper has been submitted to the GenBank/EMBL Data Bank with accession number(s) M63484.
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Issue or Number:18
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Official Citation:Cloning and functional analysis of the ubiquitin-specific protease gene UBP1 of Saccharomyces cerevisiae Tobias, J.W. et al. Journal of Biological Chemistry, Volume 266, Issue 18, 12021 - 12028
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:107998
Deposited By: George Porter
Deposited On:10 Feb 2021 23:41
Last Modified:16 Nov 2021 19:08

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