Alteration of the phospholipid composition of Escherichia coli through genetic manipulation.
In order to study the function of individual phospholipids, we have constructed a strain of Escherichia coli in which the ratio of phosphatidylethanolamine to phosphatidylglycerol plus cardiolipin can be regulated. In this strain (HDL1001) the normal expression of the phosphatidylglycerophosphate synthase does not occur due to the presence of the pgsA30 allele (Heacock, P. N., and Dowhan, W. (1987) J. Biol. Chem. 262, 13044-13049). A second chromosomal copy of the pgsA gene is fused to the lacOP region in single copy within the lac operon. Strain HDL1001 is absolutely dependent for growth on an inducer of the lac operon. In addition, the level of the pgsA gene product, the content of the two major acidic phospholipids, and the growth rate are dependent on the level of inducer in the growth medium. Cells remain viable in the absence of inducer as evidenced by a rapid return to normal growth after the readdition of inducer. The growth rate and phospholipid composition are affected only after the level of phosphatidylglycerophosphate synthase drops below about 15% of normal levels; both phosphatidic acid and (d)CDP-diacylglycerol also begin to increase to significant levels. At the point of cell arrest the level of the major acidic phospholipids is reduced by about 90% of wild type levels.
Heacock
P N
PN
Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston 77225.
Dowhan
W
W
D015139:Blotting, Southern; D002308:Cardiolipins / Q000096:biosynthesis; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D007763:Lac Operon; D009693:Nucleic Acid Hybridization; D010715:Phosphatidylglycerols / Q000096:biosynthesis; D010743:Phospholipids / Q000235:genetics / Q000378:metabolism*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism
D002308:Cardiolipins; D003470:Culture Media; D010715:Phosphatidylglycerols; D010743:Phospholipids; D007544:Isopropyl Thiogalactoside; D010744:Phosphoric Monoester Hydrolases; C026991:phosphatidylglycerophosphatase