ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/069/SRR10987369/SRR10987369.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/065/SRR10987365/SRR10987365.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/072/SRR10987372/SRR10987372.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/068/SRR10987368/SRR10987368.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/066/SRR10987366/SRR10987366.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/071/SRR10987371/SRR10987371.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/070/SRR10987370/SRR10987370.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/067/SRR10987367/SRR10987367.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/073/SRR10987373/SRR10987373.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR109/074/SRR10987374/SRR10987374.fastq.gzprimaryOK200GenomicsNational Cancer Institutehttps://www.ebi.ac.uk/ena/browser/view/PRJNA603835xref:EuropePMC:PMC9112975xref:EuropePMC:PMC7474616xref:EuropePMC:PMC9226507Bacterial transcription is punctuated by RNA polymerase (RNAP) pausing that can provide time for regulatory events to occur. However, the genome-wide role of transcription elongation factors on pausing had not been examined in vivo. We determined the genome-wide effect of Bacillus subtilis NusG on RNAP pausing using RNET-seq, a nascent RNA sequencing protocol. NusG shifts RNAP to the post-translocation register, and induces pausing at ~1600 sites containing a conserved TTNTTT motif in the non-template DNA strand within the paused transcription bubble. NusG-dependent pausing was confirmed for all pause sites tested in vitro. NusG-dependent pausing in the ribD riboswitch provides time for cotranscriptional FMN binding, which decreases the concentration of FMN required for termination.ENABacillus subtilis var. natto, Bacillus natto, Vibrio subtilis, Bacillus mesentericus, bacterial transcription, Bacillus subtilis subsp. natto, Bacillus subtilis8, Bacillus uniflagellatus., Natto Bacteria, Bacillus globigii, transcription from bacterial-type RNA polymerase promoter, Bacillus subtilis/Bacillus globigii, Bacillus subtilis (natto)falseNusG-dependent transcription pausing in Bacillus subtilis2023-05-192020-08-06PRJNA603835