1887

Abstract

In the satellite phage P4, transcription starting from P is prevalently controlled via premature termination at several termination sites. We identified a spontaneous mutation, P4 (ppression of ermination), in the natural stop codon of P4 that, by elongating translation, suppresses transcription termination at the downstream site. Both the translational and the transcriptional profile of P4 differed from those of P4 wild-type. First of all, P4 did not express Orf151, but a higher molecular mass protein, compatible with the 303 codon open reading frame generated by the fusion of , and the intervening 138 nt. Moreover, after infection of , the mutant expressed a very low amount of the 1.3 and 1.7 kb transcripts originating at P and P promoters, respectively, and terminating at the intracistronic site, whereas correspondingly higher amounts of the 4.1 and 4.5 kb RNAs arising from the same promoters and covering the entire operon were detected. Thus the mutation converts a natural stop codon into a sense codon, suppresses a natural intracistronic termination site and leads to overexpression of the downstream and genes. This correlates with the inability of P4 to propagate in the plasmid state. By cloning different P4 DNA fragments, we mapped the transcription termination site within the 7633–7361 region between and gene . A potential stem–loop structure, resembling the structure of a Rho-independent termination site, was predicted by mfold sequence analysis at 7414–7385.

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2007-03-01
2019-11-22
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