The XylS-dependent Pm promoter is transcribed in vivo by RNA polymerase with sigma 32 or sigma 38 depending on the growth phase.
The Pm promoter of the TOL plasmid of Pseudomonas putida is expressed at high level along the growth curve. This transcription is dependent on the positive regulator XylS activated by 3-methylbenzoate. The sigma factor sigma 38 is required for expression in early stationary phase and thereafter. To test whether sigma 70 was involved in Pm transcription in exponential phase, we have followed mRNA synthesis in a rpoD thermosensitive strain. No difference in Pm transcription was found between the wild type and the thermosensitive strain at the restrictive temperature of 42 degrees C, indicating that transcription was independent of the sigma factor sigma 70. However, basal levels of mRNA expression from Pm in this strain in exponential phase were more than twofold higher at 42 degrees C, suggesting involvement of sigma 32 in Pm transcription. In a rpoH background, no expression of Pm took place in the exponential phase, whereas it increased during stationary phase, and in a rpoH rpoS double mutant no activity from the Pm promoter was detected along the growth curve. We have shown that the increase in the amount of sigma 32 factor necessary for transcription in exponential phase is provided through induction of the heat shock response by the presence of the effector 3-methylbenzoate, which is also required for activation of the positive regulator XylS. We conclude that activation of Pm transcription is achieved through a switch between two stress-responsive factors, sigma 32 in exponential phase and sigma 38 in stationary phase. In both cases, transcription is dependent on the activator XylS and presents the same transcription start point.
Marqués
S
S
Consejo Superior de Investigaciones Científicas, Estación Experimental del Zaidín, Department of Plant Biochemistry, Granada, Spain.
Manzanera
M
M
González-Pérez
M M
MM
Gallegos
M T
MT
Ramos
J L
JL
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001565:Benzoates / Q000378:metabolism; D004268:DNA-Binding Proteins; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D018869:Heat-Shock Response; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016958:Pseudomonas putida / Q000235:genetics* / Q000254:growth & development; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics*
D001426:Bacterial Proteins; D001565:Benzoates; D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; D012808:Sigma Factor; D015534:Trans-Activators; D014157:Transcription Factors; C078720:XylS protein, Pseudomonas putida; C069774:heat-shock sigma factor 32; C069567:sigma factor KatF protein, Bacteria; C081775:RNA polymerase sigma 70; D012321:DNA-Directed RNA Polymerases