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PMID: 3311726 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Biochemical properties of the ras-related YPT protein in yeast: a mutational analysis.

The EMBO journal ·Vol. 6 ·No. 8 ·1987-08-00 ·Pages 2373-9

Wagner P, Molenaar CM, Rauh AJ, Brökel R, Schmitt HD, Gallwitz D

Abstract

Using site-directed mutagenesis, the ras-related and essential yeast YPT1 gene was changed to generate proteins with amino acid exchanges within conserved regions. Bacterially produced wild-type proteins were used for biochemical studies in vitro and were found to have properties very similar to mammalian ras proteins. Gene replacement allowed the study of physiological consequences of the mutations in yeast cells. Lys21----Met and Asn121----Ile substitutions rendered the protein incapable of binding GTP and caused lethality. Ser17----Gly and Ala65----Thr substitutions slightly changed the protein's apparent binding capacity for either GDP or GTP and altered its intrinsic GTPase activity. These mutations were without effect on cellular growth. The YPTgly17,thr65 mutant protein displayed a significantly altered relative capacity for guanine nucleotide binding but a GTPase activity comparable to the wild-type protein. In contrast to the Ala65----Thr substitution, the double mutant displayed a significantly reduced capacity for autophosphorylation and allowed cells to grow only poorly. Cellular growth was improved when this mutant protein was overproduced.

MeSH Terms
Amino Acid Sequence Base Sequence Fungal Proteins/genetics,metabolism Genes Genes, Fungal Guanosine Diphosphate/metabolism Guanosine Triphosphate/metabolism Kinetics Mutation Phosphorylation Saccharomyces cerevisiae/genetics ras Proteins
Chemicals
Fungal Proteins Guanosine Diphosphate Guanosine Triphosphate ras Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wagner P
Max-Planck-Institut für biophysikalische Chemie, Abteilung Molekulare Genetik, Göttingen, FRG.
Molenaar C M
Rauh A J
Brökel R
Schmitt H D
Gallwitz D
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1987-08-00
Pages
2373-9
Language
English
Region
England
NLM ID
8208664
PMCID
PMC553642
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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