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Physiologic and Transcriptomic Effects Triggered by Overexpression of Wild Type and Mutant DNA Topoisomerase I in Streptococcus pneumoniae

ABSTRACT: Physiologic and Transcriptomic Effects Triggered by Overexpression of Wild Type and Mutant DNA Topoisomerase I in Streptococcus pneumoniae

PROVIDER: PRJNA1016265 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26048963.fastq.gz	Fastqsanger.gz
	SRR26048964.fastq.gz	Fastqsanger.gz
	SRR26048965.fastq.gz	Fastqsanger.gz
	SRR26048966.fastq.gz	Fastqsanger.gz
	SRR26048967.fastq.gz	Fastqsanger.gz

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Physiologic and Transcriptomic Effects Triggered by Overexpression of Wild Type and Mutant DNA Topoisomerase I in Streptococcus pneumoniae

Project description:We obtained the topARA mutant that encodes a TopoI enzyme that shows reduced topoisomerase activity compared to the wild type gene (topAWT) and confers resistance to the antibiotic seconeolitsine. Alterations in the gene expression profile produced after overexpression of the wild type or the RA mutant of topA gene were studied. In addition, differential gene expression produced by the DNA gyrase inhibitor novobiocin (NOV) was also studied in cultures overexpressing each of the versions of TopA.

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Physiologic and Transcriptomic Effects Triggered by Overexpression of Wild Type and Mutant DNA Topoisomerase I in <i>Streptococcus pneumoniae</i>.

Project description:Topoisomerase I (TopoI) in Streptococcus pneumoniae, encoded by topA, is a suitable target for drug development. Seconeolitsine (SCN) is a new antibiotic that specifically blocks this enzyme. We obtained the topARA mutant, which encodes an enzyme less active than the wild type (topAWT) and more resistant to SCN inhibition. Likely due to the essentiality of TopoI, we were unable to replace the topAWT allele by the mutant topARA version. We compared the in vivo activity of TopoIRA and TopoIWT using regulated overexpression strains, whose genes were either under the control of a moderately (PZn) or a highly active promoter (PMal). Overproduction of TopoIRA impaired growth, increased SCN resistance and, in the presence of the gyrase inhibitor novobiocin (NOV), caused lower relaxation than TopoIWT. Differential transcriptomes were observed when the topAWT and topARA expression levels were increased about 5-fold. However, higher increases (10-15 times), produced a similar transcriptome, affecting about 52% of the genome, and correlating with a high DNA relaxation level with most responsive genes locating in topological domains. These results confirmed that TopoI is indeed the target of SCN in S. pneumoniae and show the important role of TopoI in global transcription, supporting its suitability as an antibiotic target.

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