Transcription of exogenous and endogenous deoxyribonucleic acid templates in cold-shocked Bacillus subtilis.
Ribonucleic acid (RNA) synthesis was examined in cold-shocked Bacillus subtilis cells. The cells were grown to mid-log stage, harvested, and cold shocked. RNA synthesis was monitored by the incorporation of [3H]uridine triphosphate or [alpha 32P]adenosine triphosphate into trichloroacetic acid-precipitable material in the presence of all four nucleoside triphosphates. The inhibition of RNA synthesis in cold-shocked cells by lipiarmycin, ethidium bromide, rifampin. or streptolydigin was analyzed using mutant or wild-type cells. Also examined were the effects of temperature, salt concentration, and the addition of polyamines or highly phosphorylated nucleotides. In ultraviolet-irradiated and cold-shocked cells, RNA wynthesis decreased to low levels. The addition of exogenous phi 29 or TSP-1 template to these cells caused a 13- to 20-fold increase in RNA synthesis, as monitored by trichloroacetic acid-precipitable counts. RNA synthesized in the presence of phi 29 deoxyribonucleic acid (DNA) hybridizes mainly to EcoRI fragments A and C of phi 29 DBA, These two fragments direct transcription by purified RNA polymerase in vitro and hybridize to early phi 29 DNA produced in vivo. Our results with TSP-1 DNA in this system indicated that the RNA produced hybridizes to the same fragments as early RNA produced in vivo. Plasmic pUB110 DNA was not transcribed in this system.
D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D003080:Cold Temperature; D004247:DNA / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D004996:Ethidium / Q000494:pharmacology; D009711:Nucleotides / Q000494:pharmacology; D011073:Polyamines / Q000494:pharmacology; D012329:RNA, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects
D004279:DNA, Viral; D009711:Nucleotides; D011073:Polyamines; D012329:RNA, Bacterial; D004247:DNA; D004996:Ethidium