Cloning, sequencing and chromosomal assignment of a gene from Saccharomyces cerevisiae which is negatively regulated by glucose and positively by lipids

R. L. Stone, V. Matarese, B. B. Magee, P. T. Magee, D. A. Bernlohr

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30 Scopus citations

Abstract

We report the molecular cloning, nucleotide (nt) sequence and chromosomal assignment of the Saccharomyces cerevisiae gene GLP1. This gene encoded a 15-kDa protein that was synthesized at a low level during growth on glucose and was induced ninefold upon glucose deprivation. When glucose withdrawal was accompanied by the addition of fatty acids the induction was enhanced an additional two-to threefold. The GLP1 gene product was identified as a soluble protein and purified using a combination of gel permeation and ion exchange chromatography. Using oligodeoxyribonucleotides as hybridization probes we have isolated the GLP1 gene and sequenced the single, long open reading frame which is 351 nt in length and is not interrupted by introns. The GLP1 gene directed the transcription of a 700-nt mRNA in response to glucose deprivation. The accumulation of the mRNA was further enhanced twofold by the addition of oleate. We have localized the GLP1 gene to S. cerevisiae chromosome VI.

Original languageEnglish (US)
Pages (from-to)171-176
Number of pages6
JournalGene
Volume96
Issue number2
DOIs
StatePublished - Dec 15 1990

Bibliographical note

Funding Information:
We thank Ron Scamurra for his assistance in yeast growth and John Ziemer for his assistance with CHEF. Also, we would like to thank Dr. Teresa Dunn for kindly providing the yeast actin eDNA clone. This work was supported by NIH Training Grant 5T32-GM07323 to V.M. and NIH Grant GM-43199 to D.A.B. The nt sequence data reported in this paper have been submitted to GenBank and assigned accession number M37774.

Keywords

  • CHEF pulsed-field electrophoresis
  • Recombinant DNA
  • glucose deprivation
  • oleate
  • protein purification

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