Influence of the GCGC discriminator motif introduced into the ribosomal RNA P2- and tac promoter on growth-rate control and stringent sensitivity - PubMed (original) (raw)

Influence of the GCGC discriminator motif introduced into the ribosomal RNA P2- and tac promoter on growth-rate control and stringent sensitivity

M Zacharias et al. EMBO J. 1989 Nov.

Abstract

The synthesis of stable RNA in bacteria is known to be regulated by a stringent control mechanism. Characteristic of stringent-regulated promoters, all ribosomal RNA promoters P1, but not P2, contain a GC-rich discriminator sequence assumed to be important for such a control. Using site-directed mutagenesis we have altered both the rrnB P2 and the synthetic tac promoter to the consensus GCGC discriminator motif. The modified promoters were placed upstream of the structural gene encoding the chloramphenicol acetyltransferase. The response of the modified promoters to amino acid starvation, changes in the growth rate or differences in the basal level of guanosine tetraphosphate (ppGpp) were determined in vivo. The results clearly show, that the discriminator motif is sufficient to convert the ribosomal RNA promoter P2 to a stringent, as well as growth-rate regulated, promoter. By contrast, the same discriminator sequence linked to the synthetic tac promoter does not convert this promoter to either stringency or growth-rate regulation. Finally, the results presented in this study reinforce the view that stringent and growth-rate regulation utilize the same mechanism, with ppGpp being the common mediator.

PubMed Disclaimer

Cited by

The signal for growth rate control and stringent sensitivity in E. coli is not restricted to a particular sequence motif within the promoter region.
Zacharias M, Göringer HU, Wagner R. Zacharias M, et al. Nucleic Acids Res. 1990 Nov 11;18(21):6271-5. doi: 10.1093/nar/18.21.6271. Nucleic Acids Res. 1990. PMID: 2243774 Free PMC article.
Localization of the intrinsically bent DNA region upstream of the E.coli rrnB P1 promoter.
Gaal T, Rao L, Estrem ST, Yang J, Wartell RM, Gourse RL. Gaal T, et al. Nucleic Acids Res. 1994 Jun 25;22(12):2344-50. doi: 10.1093/nar/22.12.2344. Nucleic Acids Res. 1994. PMID: 8036162 Free PMC article.
A proximal promoter element required for positive transcriptional control by guanosine tetraphosphate and DksA protein during the stringent response.
Gummesson B, Lovmar M, Nyström T. Gummesson B, et al. J Biol Chem. 2013 Jul 19;288(29):21055-21064. doi: 10.1074/jbc.M113.479998. Epub 2013 Jun 7. J Biol Chem. 2013. PMID: 23749992 Free PMC article.
The architecture and ppGpp-dependent expression of the primary transcriptome of Salmonella Typhimurium during invasion gene expression.
Ramachandran VK, Shearer N, Jacob JJ, Sharma CM, Thompson A. Ramachandran VK, et al. BMC Genomics. 2012 Jan 17;13:25. doi: 10.1186/1471-2164-13-25. BMC Genomics. 2012. PMID: 22251276 Free PMC article.
Physiological effects of the fructose-1,6-diphosphate aldolase ts8 mutation on stable RNA synthesis in Escherichia coli.
Singer M, Walter WA, Cali BM, Rouviere P, Liebke HH, Gourse RL, Gross CA. Singer M, et al. J Bacteriol. 1991 Oct;173(19):6249-57. doi: 10.1128/jb.173.19.6249-6257.1991. J Bacteriol. 1991. PMID: 1717436 Free PMC article.

References

1. Mol Gen Genet. 1984;195(3):391-401 - PubMed
1. Cell. 1985 Feb;40(2):319-26 - PubMed
1. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92 - PubMed
1. Cell. 1985 May;41(1):6-8 - PubMed
1. EMBO J. 1985 Apr;4(4):1053-8 - PubMed

MeSH terms

Substances

LinkOut - more resources

Full Text Sources
Miscellaneous
- NCI CPTAC Assay Portal

Influence of the GCGC discriminator motif introduced into the ribosomal RNA P2- and tac promoter on growth-rate control and stringent sensitivity - PubMed (original) (raw)