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Regulation of phospholipid biosynthesis in Escherichia coli. Cloning of the structural gene for the biosynthetic sn-glycerol-3-phosphate dehydrogenase.

Publication ,  Journal Article
Clark, D; Lightner, V; Edgar, R; Modrich, P; Cronan, JE; Bell, RM
Published in: J Biol Chem
January 25, 1980

The structural gene for the Escherichia coli biosynthetic sn-glycerol-3-phosphate (glycerol-P) dehydrogenase gpsA, was transferred from a defective transducing phage (lambda dcysE, gpsA) into the Eco RI site of plasmid pMB9 by recombinant DNA techniques. The recombinant plasmids suppressed the glycerol-P requirement of gpsA- mutants and strains bearing one such plasmid, pDC2, overproduced the glycerol-P dehydrogenase about 60-fold. The glycerol-P dehydrogenase from a strain bearing the pDC2 was purified 200-fold to homogeneity. This is contrasted to the 12,000-fold purification required to purify the enzyme from a wild type strain (Edgar, J. R., and Bell, R. M. (1978) J. Biol. Chem. 253, 6348-6353). The homogeneous enzyme purified from a strain bearing the pDC2 plasmide was strongly inhibited by glycerol-P (Ki of 2.5 microM). The introduction of the pDC2 plasmid into glycerol-P auxotrophs containing a Km-defective glycerol-P acyltranferase, defined by the plsB locus, caused a 60-fold overproduction of the glycerol-P requirement. This strongly suggests that the intracellular level of glycerol-P is stringently regulated in vivo by a mechanism involving feedback inhibition of the glycerol-P dehydrogenase by glycerol-P.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

January 25, 1980

Volume

255

Issue

2

Start / End Page

714 / 717

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Species Specificity
  • Protein Biosynthesis
  • Phospholipids
  • Mutation
  • Glycerolphosphate Dehydrogenase
  • Genotype
  • Genes
  • Escherichia coli
  • DNA, Recombinant
 

Citation

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Clark, D., Lightner, V., Edgar, R., Modrich, P., Cronan, J. E., & Bell, R. M. (1980). Regulation of phospholipid biosynthesis in Escherichia coli. Cloning of the structural gene for the biosynthetic sn-glycerol-3-phosphate dehydrogenase. J Biol Chem, 255(2), 714–717.
Clark, D., V. Lightner, R. Edgar, P. Modrich, J. E. Cronan, and R. M. Bell. “Regulation of phospholipid biosynthesis in Escherichia coli. Cloning of the structural gene for the biosynthetic sn-glycerol-3-phosphate dehydrogenase.J Biol Chem 255, no. 2 (January 25, 1980): 714–17.
Clark D, Lightner V, Edgar R, Modrich P, Cronan JE, Bell RM. Regulation of phospholipid biosynthesis in Escherichia coli. Cloning of the structural gene for the biosynthetic sn-glycerol-3-phosphate dehydrogenase. J Biol Chem. 1980 Jan 25;255(2):714–7.
Clark D, Lightner V, Edgar R, Modrich P, Cronan JE, Bell RM. Regulation of phospholipid biosynthesis in Escherichia coli. Cloning of the structural gene for the biosynthetic sn-glycerol-3-phosphate dehydrogenase. J Biol Chem. 1980 Jan 25;255(2):714–717.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

January 25, 1980

Volume

255

Issue

2

Start / End Page

714 / 717

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Species Specificity
  • Protein Biosynthesis
  • Phospholipids
  • Mutation
  • Glycerolphosphate Dehydrogenase
  • Genotype
  • Genes
  • Escherichia coli
  • DNA, Recombinant