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Heat-Inducible Degron: A Method for Constructing Temperature-Sensitive Mutants

Dohmen, R. Jürgen and Wu, Peipei and Varshavsky, Alexander (1994) Heat-Inducible Degron: A Method for Constructing Temperature-Sensitive Mutants. Science, 263 (5151). pp. 1273-1276. ISSN 0036-8075. https://resolver.caltech.edu/CaltechAUTHORS:20150120-134752646

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Abstract

A temperature-sensitive (ts) mutant retains the function of a gene at a low (permissive) temperature but not at a high (nonpermissive) temperature. Arg-DHFR, a dihydrofolate reductase bearing an amino-terminal (N-terminal) arginine, is long-lived in the yeast Saccharomyces cerevisiae, even though arginine is a destabilizing residue in the N-end rule of protein degradation. A ts derivative of Arg-DHFR was identified that is long-lived at 23°C but rapidly degraded by the N-end rule pathway at 37°C. Fusions of ts Arg-DHFR to either Ura3 or Cdc28 of S. cerevisiae confer ts phenotypes specific for these gene products. Thus, Arg-DHFR^ts is a heat-inducible degradation signal that can be used to produce ts mutants without a search for ts mutations.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1126/science.8122109 DOIArticle
http://www.jstor.org/stable/2883471JSTORArticle
ORCID:
AuthorORCID
Varshavsky, Alexander0000-0002-4011-258X
Additional Information:© 1994 American Association for the Advancement of Science. 27 September 1993; accepted 21 January 1994. We thank N. Kleckner (Harvard University) for the gift of plasmids and members of this laboratory, especially E. Johnson, K. Madura, F. Lévy, C. Byrd, and T. Clandinin, for plasmids and strains and for advice and comments on the manuscript. Supported by grants to A.V. from the National Institutes of Health. R.J.D. was supported by a postdoctoral fellowship from the European Molecular Biology Organization.
Funders:
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NIHUNSPECIFIED
European Molecular Biology Organization (EMBO)UNSPECIFIED
Issue or Number:5151
Record Number:CaltechAUTHORS:20150120-134752646
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150120-134752646
Official Citation:Dohmen, R., Wu, P., & Varshavsky, A. (1994). Heat-inducible degron: a method for constructing temperature-sensitive mutants. Science, 263(5151), 1273-1276. doi: 10.1126/science.8122109
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:53889
Collection:CaltechAUTHORS
Deposited By: Jason Perez
Deposited On:21 Jan 2015 18:25
Last Modified:25 Nov 2020 22:53

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