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Codon-usage based regulation of colicin K synthesis by the stress alarmone ppGpp by Irena Kuhar; Jos P. M. Van Putten; Darja Žgur-Bertok; Wim Gaastra; Bart J. A. M. Jordi is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Codon-usage based regulation of colicin K synthesis by the stress alarmone ppGpp about?

The molecular mechanism of the upregulation of __Escherichia coli__ colicin K (Cka) synthesis during stress conditions was studied. Nutrient starvation experiments and the use of __relA spoT__ mutant strains, IPTG‐regulated overproduction of ppGpp and __lacZ__ fusions revealed that the stringent response alarmone guanosine 3′,5′‐bispyrophosphate (ppGpp) is the main positive effector of Cka synthesis. Comparison of the amounts of protein produced (Western blotting) and specific mRNA (Northern blotting) before and after nutrient starvation demonstrated increases in Cka protein with unaltered specific mRNA levels, suggesting a post‐transcriptional regulatory mechanism. Reporter (β‐galactosidase) assays using truncated __cka__ of variable length fused to __lacZ__ located the key regulatory region close to the 5′ end of the __cka__ mRNA. Closer analysis of this region indicated the presence of several rare codons, including the leucine‐encoding codon CUA. Synonymous exchange of the rare codons with more frequently used ones abolished the regulatory effect of ppGpp. Supplementation of the strain with the plasmid CodonPlus carrying several rare tRNA genes yielded similar results, indicati

Who reads Codon-usage based regulation of colicin K synthesis by the stress alarmone ppGpp?

Author

Irena Kuhar; Jos P. M. Van Putten; Darja Žgur-Bertok; Wim Gaastra; Bart J. A. M. Jordi

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0950-382X)

Published

2001

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)