Overexpression of the cat-86 gene is associated with thermosensitivity in Bacillus subtilis.
Bacillus subtilis harboring the cat-86 constitutive plasmid pPL708C2 with an ochre mutation at the 9th codon (terc 9) was sensitive to chloramphenicol (Cm(s)) and exhibited relative thermostability when heated at 47 degrees C. Reversion to chloramphenicol resistance (Cm(r)) occurred at a frequency of 5.4 x 10(-8). All of the plasmid Cm(r) revertants tested were thermosensitive. Similarly, wild-type pPL708C2 present in B. subtilis also rendered the bacterium thermosensitive. When a nonsense mutation is introduced at codon 141, however, this terc 141 variant of pPL708C2 failed to thermosensitize B. subtilis. Another variant of pPL708C2 that produces intact yet catalytically inactive CAT-86 has both His-16 and His-17 at the active site replaced by Pro. Nevertheless, cells of B. subtilis carrying this variant were thermosensitive. Plasmid-free and pPL708C2-bearing strains did not exhibit differences in major heat shock proteins. Electron micrographs revealed a threefold increase of inclusion bodies present in a strain harboring pPL708C2 when compared with those in an isogenic plasmid-free strain.
Friedman
S M
SM
Department of Biological Sciences, Hunter College of The City University of New York, 695 Park Avenue, New York, New York 10021, USA.
Chatakondu
S
S
Urbanski
D
D
Lopes
R
R
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D002701:Chloramphenicol / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D008854:Microscopy, Electron
D002701:Chloramphenicol; D015500:Chloramphenicol O-Acetyltransferase