1887

Abstract

The DNA region of the chromosome containing the heat-shock genes and was cloned, and the genes were sequenced. These genes are arranged in the chromosome in the order . Northern hybridization experiments with RNA from and a DNA probe carrying both and genes showed a single, heat-inducible transcript of approximately 2200 nt, indicating that these genes form an operon. Primer extension analysis revealed a strong, heat-inducible transcription start site 59 nt upstream of , preceded by a sequence typical for the heat-shock promoters recognized by the factor, and a weak transcription start site 25 nt upstream the gene, preceded by a sequence typical for promoters. Transcription from the latter promoter occurred only at low temperatures. The operon cloned in a plasmid in cells was transcribed in a -dependent manner; the transcript size and the -dependent transcription start site were as in cells. Comparison of transcription regulation with the other well-characterized operons of the subdivision of proteobacteria (those of and ) indicates a high conservation of the transcriptional regulatory elements among these bacteria, with two promoters, and , involved in the regulation. The ability of the cloned genes to complement mutants was tested: restored a thermoresistant phenotype to bacteria and enabled phage to grow in the mutant cells. did not abolish thermosensitivity of bacteria but it complemented the mutant with respect to growth of phage. The results suggest that the GroEL chaperone may be more species-specific than the GroES co-chaperone.

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2003-06-01
2020-01-29
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