RESUMO
Induction of stress proteins in response to heat shock was found to be reduced significantly in Escherichia coli with DeltatopA mutation. RNase H overexpression in the DeltatopA mutant partially restored the sigma(32)-dependent induction of stress genes in response to high temperature and ethanol. The presence of overexpressed RNase H also improved the survival rate of the DeltatopA mutant after high temperature and oxidative challenges. Topoisomerase I is likely required during stress response for preventing accumulation of transcription-driven hypernegative supercoiling and R-loop formation at induced stress genes loci.
Assuntos
DNA Topoisomerases Tipo I/genética , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Proteínas de Choque Térmico/biossíntese , Ribonuclease H/biossíntese , DNA Bacteriano/análise , Escherichia coli/enzimologia , Escherichia coli/crescimento & desenvolvimento , Mutação , Conformação de Ácido Nucleico , Temperatura , Transcrição GênicaRESUMO
Escherichia coli DNA topoisomerase I (encoded by the topA gene) is important for maintaining steady-state DNA supercoiling and has been shown to influence vital cellular processes including transcription. Topoisomerase I activity is also needed to remove hypernegative supercoiling generated on the DNA template by the progressing RNA polymerase complex during transcription elongation. The accumulation of hypernegative supercoiling in the absence of topoisomerase I can lead to R-loop formation by the nascent transcript and template strand, leading to suppression of transcription elongation. Here we show by affinity chromatography and overlay blotting that E. coli DNA topoisomerase I interacts directly with the RNA polymerase complex. The protein-protein interaction involves the beta' subunit of RNA polymerase and the C-terminal domains of E. coli DNA topoisomerase I, which are homologous to the zinc ribbon domains in a number of transcription factors. This direct interaction can bring the topoisomerase I relaxing activity to the site of transcription where its activity is needed. The zinc ribbon C-terminal domains of other type IA topoisomerases, including mammalian topoisomerase III, may also help link the enzyme activities to their physiological functions, potentially including replication, transcription, recombination, and repair.