Response of RNA polymerase to ppGpp: requirement for the omega subunit and relief of this requirement by DksA.
Previous studies have come to conflicting conclusions about the requirement for the omega subunit of RNA polymerase in bacterial transcription regulation. We demonstrate here that purified RNAP lacking omega does not respond in vitro to the effector of the stringent response, ppGpp. DksA, a transcription factor that works in concert with ppGpp to regulate rRNA expression in vivo and in vitro, fully rescues the ppGpp-unresponsiveness of RNAP lacking omega, likely explaining why strains lacking omega display a stringent response in vivo. These results demonstrate that omega plays a role in RNAP function (in addition to its previously reported role in RNAP assembly) and highlight the importance of inclusion of omega in RNAP purification protocols. Furthermore, these results suggest that either one or both of two short segments in the beta' subunit that physically link omega to the ppGpp-binding region of the enzyme may play crucial roles in ppGpp and DksA function.
Vrentas
Catherine E
CE
Department of Bacteriology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Gaal
Tamas
T
Ross
Wilma
W
Ebright
Richard H
RH
Gourse
Richard L
RL
D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D006159:Guanosine Tetraphosphate / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D020836:Protein Structure, Quaternary; D012335:RNA, Ribosomal / Q000096:biosynthesis* / Q000737:chemistry; D012808:Sigma Factor / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic / Q000502:physiology; D012407:rRNA Operon / Q000502:physiology
D029968:Escherichia coli Proteins; D012335:RNA, Ribosomal; D012808:Sigma Factor; C090751:dksA protein, E coli; D006159:Guanosine Tetraphosphate; C111232:RNA polymerase omega subunit; D012321:DNA-Directed RNA Polymerases