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PMID: 9753749 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Silent mutations in the Escherichia coli ompA leader peptide region strongly affect transcription and translation in vivo.

Nucleic acids research ·Vol. 26 ·No. 20 ·1998-10-15 ·Pages 4778-82

Deana A, Ehrlich R, Reiss C

Abstract

In order to test the effect of silent mutations on the regulation of gene expression, we monitored several steps of transcription and translation of the ompA gene in vivo , in which some or all codons between codons 6 and 14, frequently used in Escherichia coli , had been exchanged for infrequent synonymous codons. Northern blot analysis revealed an up to 4-fold reduction in the half-life of the mutated messengers and a >10-fold reduction in their steady-state amounts. Western blot analysis showed a 10-fold reduction in the amount of OmpA protein. Use of a system expressing a Rho-specific anti-terminator allowed us to detect a strong transcription polarity effect in the silent mutants. These results demonstrate that silent mutations can severely inhibit several steps of gene expression in E. coli and that code degeneracy is efficiently exploited in this species for setting signals for gene control and regulation.

MeSH Terms
Bacterial Outer Membrane Proteins/genetics Blotting, Northern Capsid/pharmacology Capsid Proteins Codon/genetics Escherichia coli/genetics Gene Expression Half-Life Molecular Sequence Data Mutagenesis, Site-Directed Point Mutation/genetics Protein Biosynthesis/genetics Protein Sorting Signals/genetics RNA, Antisense RNA, Messenger/metabolism Temperature Transcription, Genetic/genetics
Chemicals
Bacterial Outer Membrane Proteins Capsid Proteins Codon Protein Sorting Signals RNA, Antisense RNA, Messenger polarity suppression factor, bacteriophage P4 OMPA outer membrane proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Deana A
Centre de Génétique Moléculaire, Laboratoire Structure et Dynamique du Génome, CNRS, F91198 Gif-sur-Yvette, France. [email protected]
Ehrlich R
Reiss C
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1998-10-15
Pages
4778-82
Language
English
Region
England
NLM ID
0411011
PMCID
PMC147888
Subset
IM
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