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Isolation of the fructose-1,6-bisphosphatase gene of the yeast Schizosaccharomyces pombe. Evidence for transcriptional regulation.

Authors
Type
Published Article
Journal
The Journal of biological chemistry
Publication Date
Volume
260
Issue
10
Pages
6348–6353
Identifiers
PMID: 2987220
Source
Medline
License
Unknown

Abstract

The fructose-1,6-bisphosphatase structural gene (FBP+) of Schizosaccharomyces pombe has been isolated by genetic complementation of a deficient mutant, which is characterized by the inability to grow on a nonfermentable carbon source such as glycerol. Growth on glycerol-containing medium was restored in a S. pombe fructose-1,6-bisphosphatase-deficient mutant (fbp-16) when it was transformed with a plasmid (pAVO4) carrying FBP+. The transformant displayed a 5-fold increase in enzymatic activity when compared to the parental S. pombe strain. Subcloning of DNA fragments from the 8.5-kilobase (kb) insert of pAVO4 defined a 4-kb DNA fragment which contained the functional FBP+ gene and its regulatory region. When this gene was placed under the control of the lac promoter-operator, functional expression in Escherichia coli was obtained, as deduced by complementation of bacterial fructose-1,6-bisphosphatase mutants. The FBP+ gene encodes a 1.9-kb glucose-repressible transcript whose appearance in S. pombe is correlated with fructose-1,6-bisphosphatase derepression in glycerol-containing medium. We suggest that the regulation of the S. pombe FBP+ gene is exerted at the transcriptional level. The S. pombe FBP+ gene gave rise to a 1.9-kb transcript in Saccharomyces cerevisiae, but not to measurable enzymatic activity.

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