Document Detail


The differential effects of guanosine tetraphosphate on open complex formation at the Escherichia coli ribosomal protein promoters rplJ and rpsA P1.
MedLine Citation:
PMID:  9810685     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The effects of guanosine tetraphosphate (ppGpp) on inhibition of single-round in vitro transcription and on the kinetics of open complex formation were investigated at the Escherichia coli ribosomal protein promoters rplJ and rpsA P1. The two promoters differ in their saturation characteristics and sensitivities to ppGpp. With a 10:1 molar ratio of RNA polymerase (RNAP) to DNA, saturation of transcription activity and weak inhibition (approximately 30%) are observed at rplJ, in contrast to the weak activity and strong inhibition (approximately 80%) at rpsA P1. In the absence of ppGpp, the two promoters show a threefold difference in the overall rate constants of association (ka) (6.5 x 10(7) M-1 s-1 at rplJ and 2.0 x 10(7) M-1 s-1 at rpsA P1), while the dissociation rate constants (kd) are similar (approximately 4.8 x 10(-5) s-1). The addition of ppGpp causes a twofold reduction in k2 (isomerisation constant) rplJ and a threefold decrease in KB (equilibrium constant of RNAP binding) at rpsA P1. There is a significant twofold increase in kd at rplJ, compared with smaller changes at rpsA P1 and at the non-stringent lacUV5 promoter. These results indicate that ppGpp affects the formation and stability of the open complex at the rplJ promoter, in contrast to the inhibition of RNAP binding to the rpsA P1 promoter.
Authors:
A Raghavan; D B Kameshwari; D Chatterji
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biophysical chemistry     Volume:  75     ISSN:  0301-4622     ISO Abbreviation:  Biophys. Chem.     Publication Date:  1998 Oct 
Date Detail:
Created Date:  1998-12-09     Completed Date:  1998-12-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0403171     Medline TA:  Biophys Chem     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  7-19     Citation Subset:  IM    
Affiliation:
Centre for Cellular and Molecular Biology, Hyderabad, India.
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MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / metabolism*
Collodion
DNA Fragmentation
DNA-Directed RNA Polymerases / metabolism
Escherichia coli / drug effects,  genetics,  metabolism*
Guanosine Tetraphosphate / pharmacology*
Kinetics
Ribosomal Proteins / metabolism*
Transcription, Genetic / genetics
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/JofD protein, Bacillus subtilis; 0/Ribosomal Proteins; 0/ribosomal protein L10; 33503-72-9/Guanosine Tetraphosphate; 9004-70-0/Collodion; EC 2.7.7.6/DNA-Directed RNA Polymerases

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