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Science 4 March 1994:
Vol. 263. no. 5151, pp. 1273 - 1276
DOI: 10.1126/science.8122109
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Articles

Science, Vol 263, Issue 5151, 1273-1276
Copyright © 1994 by American Association for the Advancement of Science

articles

Heat-inducible degron: a method for constructing temperature-sensitive mutants

RJ Dohmen, P Wu, and A Varshavsky

Division of Biology, California Institute of Technology, Pasadena 91125.

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 degrees C but rapidly degraded by the N-end rule pathway at 37 degrees C. Fusions of ts Arg-DHFR to either Ura3 or Cdc28 of S. cerevisiae confer ts phenotypes specific for these gene products. Thus, Arg-DHFRts is a heat-inducible degradation signal that can be used to produce ts mutants without a search for ts mutations.


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A Yeast Ubc9 Mutant Protein with Temperature-sensitive in Vivo Function Is Subject to Conditional Proteolysis by a Ubiquitin- and Proteasome-dependent Pathway.
J. Betting and W. Seufert (1996)
J. Biol. Chem. 271, 25790-25796
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Modular Structure of Glucocorticoid Receptor Domains Is Not Equivalent to Functional Independence. STABILITY AND ACTIVITY OF THE STEROID BINDING DOMAIN ARE CONTROLLED BY SEQUENCES IN SEPARATE DOMAINS.
M. Xu, P. K. Chakraborti, M. J. Garabedian, K. R. Yamamoto, and S. S. Simons Jr. (1996)
J. Biol. Chem. 271, 21430-21438
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Multiple functions for the poly(A)-binding protein in mRNA decapping and deadenylation in yeast..
G Caponigro and R Parker (1995)
Genes & Dev. 9, 2421-2432
   Abstract »    PDF »
A Proteolytic Pathway That Recognizes Ubiquitin as a Degradation Signal.
E. S. Johnson, P. C. M. Ma, I. M. Ota, and A. Varshavsky (1995)
J. Biol. Chem. 270, 17442-17456
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Yeast N-terminal Amidase.
R. T. Baker and A. Varshavsky (1995)
J. Biol. Chem. 270, 12065-12074
   Abstract »    Full Text »    PDF »
Methotrexate Inhibits Proteolysis of Dihydrofolate Reductase by the N-end Rule Pathway.
J. A. Johnston, E. S. Johnson, P. R. H. Waller, and A. Varshavsky (1995)
J. Biol. Chem. 270, 8172-8178
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The N-end Rule.
A. Varshavsky (1995)
Cold Spring Harb Symp Quant Biol 60, 461-478
   Abstract »    PDF »
Rvb1p and Rvb2p Are Essential Components of a Chromatin Remodeling Complex That Regulates Transcription of over 5% of Yeast Genes.
Z. O. Jonsson, S. K. Dhar, G. J. Narlikar, R. Auty, N. Wagle, D. Pellman, R. E. Pratt, R. Kingston, and A. Dutta (2001)
J. Biol. Chem. 276, 16279-16288
   Abstract »    Full Text »    PDF »
Implication of a novel multiprotein Dam1p complex in outer kinetochore function.
I. M. Cheeseman, C. Brew, M. Wolyniak, A. Desai, S. Anderson, N. Muster, J. R. Yates, T. C. Huffaker, D. G. Drubin, and G. Barnes (2001)
J. Cell Biol. 155, 1137-1146
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Essential Role of MCM Proteins in Premeiotic DNA Replication.
K. Lindner, J. Gregan, S. Montgomery, and S. E. Kearsey (2002)
Mol. Biol. Cell 13, 435-444
   Abstract »    Full Text »    PDF »


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