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Genetics, Vol 119, 743-749, Copyright © 1988
INVESTIGATIONS |
Autogenous Regulation of the regA Gene of Bacteriophage T4: Derepression of Translation
Y. Liang, R. Wei, T. Hsu, C. Alford, M. Dawson and J. Karam
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425
The regA gene of phage T4 encodes a translational repressor that inhibits utilization of its own mRNA as well as the translation of a number of other phage-induced mRNAs. In recombinant plasmids, autogenous translational repression limits production of the RegA protein when the cloned structural gene is expressed under control of a strong, plasmid-borne promoter (lambda P(L)). We have found that a genetic fusion which places the regA ribosome binding domain in proximity to active translation leads to partial derepression of wild-type RegA protein synthesis. The derepression is not due to increased synthesis of regA RNA, suggesting that it occurs at the translational level. Derepressed clones of the wild-type regA gene were used to overproduce and purify the repressor. In an in vitro assay the wild-type target was sensitive and a mutant target was resistant to inhibition by the added protein. The results suggest that the sensitivity of a regA-regulated cistron to translational repression may depend on the competition between ribosomes and RegA protein for overlapping recognition sequences in the translation initiation domain of the mRNA.
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