A trans-acting regulatory mutation that causes overproduction of phosphatidylserine synthase in Escherichia coli.

We have isolated three mutants of Escherichia coli which have elevated levels of the phospholipid synthetic enzyme phosphatidylserine synthase. One of these strains carries a mutation, designated pssR1, which maps near minute 84 of the chromosome, distinct from the synthase structural gene (pss) at minute 56. The pssR1 mutation causes selective overproduction of phosphatidylserine synthase, since the levels of six other lipid synthetic enzymes are unaltered. The specific activity of the synthase in crude cell extracts of mutants harboring pssR1 is about five times greater than wild type. The synthase can also be overproduced 10-fold in wild type strains with hybrid ColE1 plasmids carrying the synthase structural gene (pss). A pssR1 mutant harboring such a pss plasmid overproduces the synthase about 50-fold. This multiplicative interaction of pssR1 and cloned pss demonstrates that pssR1 is trans-acting. The synthase has been purified in parallel from pssR1 and pssR+ strains. The pssR1 mutant yields more total synthase protein than pssR+, but the pure enzyme has the same specific activity in both cases. Therefore, pssR1 acts by increasing the amount of the normal protein, not by activating the enzyme. The discovery of pssR shows that there are regulatory loci which control the production of enzymes involved in membrane lipid synthesis.