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Design and Isolation of Temperature-sensitive Mutants of Gal4 in Yeast and Drosophila

Mondal, Kajari and Dastidar, Antara Ghosh and Singh, Guramrit and Madhusudhanan, S and Gande, Santosh Lakshmi and VijayRaghavan, K and Varadarajan, Raghavan (2007) Design and Isolation of Temperature-sensitive Mutants of Gal4 in Yeast and Drosophila. In: Journal of Molecular Biology, 370 (5). pp. 939-950.

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Little is known about mechanisms responsible for the temperature-sensitive (ts) phenotype, or of the transferability of ts mutants of a specific gene between organisms. Using a structure-based approach, nine ts mutants of Gal4 were generated in yeast by mutating four DNA binding residues. Two of these nine yeast ts mutants were cloned into P element vectors under control of the Elav and GMR promoters and transgenic Drosophila lines were generated. These were crossed to UAS reporter lines and progeny were characterized for reporter gene expression as a function of temperature. Both of these yeast ts mutants show a ts phenotype in Drosophila and result in rapid induction of reporter gene expression upon shifting to the permissive temperature. Exposed, functional residues involved in protein–ligand or protein–protein interactions appear to be attractive candidate sites for generating ts mutants that are transferable between organisms.

Item Type: Journal Article
Publication: Journal of Molecular Biology
Publisher: Elsevier
Additional Information: Copyright of this article belongs to Elsevier.
Keywords: Temperature-sensitive;Design;Transferability;Yeast;Drosophila
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited: 27 Sep 2007
Last Modified: 19 Sep 2010 04:39
URI: http://eprints.iisc.ac.in/id/eprint/11941

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