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Proc Natl Acad Sci U S A. 2016 May 3;113(18):E2506-15. doi: 10.1073/pnas.1604190113. Epub 2016 Apr 18.

Rational elicitation of cold-sensitive phenotypes.

Author information

1
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560 012, India;
2
National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore 560 065, India; vijay@ncbs.res.in varadar@mbu.iisc.ernet.in.
3
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560 012, India; Jawaharlal Center for Advanced Scientific Research, Bangalore 560 004, India vijay@ncbs.res.in varadar@mbu.iisc.ernet.in.

Abstract

Cold-sensitive phenotypes have helped us understand macromolecular assembly and biological phenomena, yet few attempts have been made to understand the basis of cold sensitivity or to elicit it by design. We report a method for rational design of cold-sensitive phenotypes. The method involves generation of partial loss-of-function mutants, at either buried or functional sites, coupled with selective overexpression strategies. The only essential input is amino acid sequence, although available structural information can be used as well. The method has been used to elicit cold-sensitive mutants of a variety of proteins, both monomeric and dimeric, and in multiple organisms, namely Escherichia coli, Saccharomyces cerevisiae, and Drosophila melanogaster This simple, yet effective technique of inducing cold sensitivity eliminates the need for complex mutations and provides a plausible molecular mechanism for eliciting cold-sensitive phenotypes.

KEYWORDS:

cold sensitivity; conditional mutants; heat-induced expression; rational design; transfer between organisms

PMID:
27091994
PMCID:
PMC4983833
DOI:
10.1073/pnas.1604190113
[Indexed for MEDLINE]
Free PMC Article

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