Osmotic Reversal of Temperature Sensitivity in Escherichia coli

Forty temperature-sensitive mutants, unable to grow on tryptone or nutrient agar at 42 C, were isolated from Escherichia coli K-12. When 0.5% NaCl was added to the medium, 32 grew at the nonpermissive temperature. Several were tested with different amounts of NaCl added to tryptone broth; all grew best when the osmolality of the medium was between 400 and 1,000 milliosmolal. One of the mutants was studied in more detail. Sucrose, inositol, KCl, and MgCl(2), as well as NaCl, permitted growth at 42 C. Glycerol, however, had no effect. When shifted from 30 to 42 C without osmotic protection, the mutant stopped growing but did not lyse, die, or leak significant amounts of intracellular material. In a similar shift experiment, a second mutant leaked all of its trichloroacetic acid-soluble pools into the medium. The majority of the mutants were hypersensitive to certain antibiotics, indicating possible cell envelope defects.