The role of the alarmone (p)ppGpp in sigma N competition for core RNA polymerase.
Some promoters, including the DmpR-controlled sigma(N)-dependent Po promoter, are effectively rendered silent in cells lacking the nutritional alarmone (p)ppGpp. Here we demonstrate that four mutations within the housekeeping sigma(D)-factor can restore sigma(N)-dependent Po transcription in the absence of (p)ppGpp. Using both in vitro and in vivo transcription competition assays, we show that all the four sigma(D) mutant proteins are defective in their ability to compete with sigma(N) for available core RNA polymerase and that the magnitude of the defect reflects the hierarchy of restoration of transcription from Po in (p)ppGpp-deficient cells. Consistently, underproduction of sigma(D) or overproduction of the anti-sigma(D) protein Rsd were also found to allow (p)ppGpp-independent transcription from the sigma(N)-Po promoter. Together with data from the direct effects of (p)ppGpp on sigma(N)-dependent Po transcription and sigma-factor competition, the results support a model in which (p)ppGpp serves as a master global regulator of transcription by differentially modulating alternative sigma-factor competition to adapt to changing cellular nutritional demands.
Laurie
Andrew D
AD
Department of Molecular Biology, Umeå University, 901 87 Umeå, Sweden.
Bernardo
Lisandro M D
LM
Sze
Chun Chau
CC
Skarfstad
Eleonore
E
Szalewska-Palasz
Agnieszka
A
Nyström
Thomas
T
Shingler
Victoria
V
D001483:Base Sequence; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D006159:Guanosine Tetraphosphate / Q000378:metabolism*; D009154:Mutation; D014158:Transcription, Genetic / Q000235:genetics
D017931:DNA Primers; D006159:Guanosine Tetraphosphate; D012321:DNA-Directed RNA Polymerases