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PMID: 331264 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Inhibition of transcription of supercoiled PM2 DNA by carbodiimide modification.

Nucleic acids research ·Vol. 4 ·No. 6 ·1977-06-00 ·Pages 1713-26

Flashner MS, Katopes MA, Lebowitz J

Abstract

PM2 superhelican DNA (form I), which as been reacted with the single strand specific reagent, N-cyclohexyl-N'-beta-(methylmorpholinium)ethyl carbodiimide (CMC) is more than 95% inhibited in its ability to support transcription with E. coli B RNA polymerase in vitro. Almost complete inhibition of transcription was achieved after 2 hours of reaction with FI when only 1% of the bases were modified. A large increase in S20,* (from 26.8 S to 33.6 S) of FI DNA was observed during the course of reaction. Rifampicin resistant transcription is more susceptible to inhibition by CMC than total transcription, suggesting that the CMC is preferentially binding at promoter sites. These results clearly are in accord with the observation that supercoiled DNA contains localized regions of unpaired bases. The promotor sites for E. coli RNA polymerase in FI PM2 DNA appear to be located at or near these unpaired sites.

MeSH Terms
Carbodiimides/pharmacology Centrifugation, Density Gradient DNA, Viral/metabolism DNA-Directed RNA Polymerases/metabolism Escherichia coli Kinetics Morpholines/pharmacology Nucleic Acid Conformation/drug effects Rifampin/pharmacology Transcription, Genetic/drug effects Ultracentrifugation
Chemicals
Carbodiimides DNA, Viral Morpholines DNA-Directed RNA Polymerases Rifampin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Flashner M S
Katopes M A
Lebowitz J
References (34)
34 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/4.6.1713
Published
1977-06-00
Pages
1713-26
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342516
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/4.6.1713 PMC Full Text View journal details
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