Document Detail

A gene encoding sn-glycerol 3-phosphate dehydrogenase (NAD+) complements an osmosensitive mutant of Saccharomyces cerevisiae.
MedLine Citation:
PMID:  7934860     Owner:  NLM     Status:  MEDLINE    
Osmoregulatory mutants of Saccharomyces cerevisiae with a defect in their capacity to readjust the cell volume/buoyant density after osmotically induced dehydration were enriched by density gradient centrifugation. Colonies derived from cells that remained dense after dehydration were screened for sensitivity to high concentrations of NaCl and defects in their osmotically induced production and intracellular accumulation of glycerol. The isolated osg (osmosensitive glycerol defective) mutants were recessive in heterozygous diploids and fell into four complementation groups (osg1-osg4). The osg1-1 mutant, described in this work, is unable to grow at low water potential and shows a decreased capacity for glycerol production and a strongly reduced activity of NAD(+)-dependent sn-glycerol 3-phosphate dehydrogenase (GPD), an enzyme in the glycerol-producing pathway. Complementation of the osg1-1 salt sensitivity defect with a low copy yeast genomic library led to the cloning of GPD1, encoding an S. cerevisiae GPD consisting of 391 amino acids and sharing 47-50% identity with GPD from other sources. Micro-sequencing of the N-terminus of purified S. cerevisiae GPD revealed a 20-amino-acid sequence that was identical to a nucleotide-deduced amino acid sequence in GPD1, but indicated that the enzyme is produced with an N-terminal extension that is removed from the functional enzyme. Subcellular fractionation does not indicate, however, that the putative pre-sequence targets GPD to any organelle; the enzyme appears to be located in the cytoplasm. Chromoblot and tetrad analysis were used to position the GPD1 gene to chromosome IV, with a distance of about 18 cM from trp1.
K Larsson; R Ansell; P Eriksson; L Adler
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular microbiology     Volume:  10     ISSN:  0950-382X     ISO Abbreviation:  Mol. Microbiol.     Publication Date:  1993 Dec 
Date Detail:
Created Date:  1994-11-23     Completed Date:  1994-11-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8712028     Medline TA:  Mol Microbiol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  1101-11     Citation Subset:  IM    
Department of General and Marine Microbiology, University of Göteborg, Sweden.
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MeSH Terms
Amino Acid Sequence
Cloning, Molecular
Genes, Fungal*
Genetic Complementation Test
Glycerol-3-Phosphate Dehydrogenase (NAD+)
Glycerolphosphate Dehydrogenase / genetics*
Molecular Sequence Data
Restriction Mapping
Saccharomyces cerevisiae / enzymology*,  genetics*
Sequence Homology, Amino Acid
Water-Electrolyte Balance / genetics
Reg. No./Substance:
EC 1.1.-/Glycerolphosphate Dehydrogenase; EC Dehydrogenase (NAD+)

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