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Cell cycle-associated expression patterns predict gene function in mycobacteria

ABSTRACT: Cell cycle-associated expression patterns predict gene function in mycobacteria

PROVIDER: PRJNA613916 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11364814_1.fastq.gz	Fastqsanger.gz
	SRR11364814_2.fastq.gz	Fastqsanger.gz
	SRR11364815_1.fastq.gz	Fastqsanger.gz
	SRR11364815_2.fastq.gz	Fastqsanger.gz
	SRR11364816_1.fastq.gz	Fastqsanger.gz

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Cell cycle-associated expression patterns predict gene function in mycobacteria

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Project description:RNA polymerase (RNAP) binding protein RbpA contributes to the formation of stable RNAP-promoter open complexes (RPo) and is essential for viability in mycobacteria. Based on structural and biochemical data, four domains have been identified in the RbpA protein: a N-terminal tail (NTT) domain of unknown function, a core domain (CD) that contacts the RNAP β’ subunit in a recently solved crystal structure, a basic linker (BL) that binds DNA, and a s-interaction domain (SID) that binds group I and group II s-factors. However, limited in vivo studies have been performed in mycobacteria and how the individual structural domains of RbpA contribute to RbpA function and mycobacterial gene expression remains mostly unknown. We dissected the roles of the RbpA structural domains in mycobacteria using a panel of rbpA mutants that target individual RbpA domains. The function of each RbpA domain was required for Mycobacterium tuberculosis viability and optimal growth in Mycobacterium smegmatis. We determined that the RbpA SID is both necessary and sufficient for RbpA interaction with the RNAP holoenzyme, indicating that the primary function of the CD is not solely association with the RNAP. We show that RbpA BL and SID are required for stabilization of RPo complexes at the ribosomal RNA rrnAP3 promoter in vitro, while the NTT and CD are dispensable. Finally, we determine that the NTT and CD impact gene expression of a distinct set of genes from that affected by the BL and SID activities. Our findings highlight specific outcomes for the activities of the individual functional domains in RbpA.

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