ENAapplication/xmlftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502963/SRR502963.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502961/SRR502961.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502959/SRR502959.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502962/SRR502962.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502964/SRR502964.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502966/SRR502966.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502960/SRR502960.fastq.gzftp.sra.ebi.ac.uk/vol1/fastq/SRR502/SRR502965/SRR502965.fastq.gzprimaryOK2000000GenomicsSteinmetz Lab, Genome Biology Unit, EMBL Heidelberghttps://www.ebi.ac.uk/ena/browser/view/PRJNA167771Saccharomyces cerevisiaeAnti-sense non-coding transcripts, genes-within-genes, and convergent gene pairs are prevalent among eukaryotes. The existence of such transcription units raises the question of what happens when RNA polymerase II (RNAPII) molecules collide head-to-head. Here we use a combination of biochemical and genetic approaches in yeast to show that polymerases transcribing opposite DNA strands cannot bypass each other. RNAPII stops, but does not dissociate upon head-to-head collision in vitro, suggesting that opposing polymerases represent insurmountable obstacles for each other. Head-to-head collision in vivo results in RNAPII stopping as well, and removal of collided RNAPII from the DNA template can be achieved via ubiquitylation-directed proteolysis. Indeed, in cells lacking efficient RNAPII poly-ubiquitylation, the half-life of collided polymerases increases, so that these can be detected between convergent genes by ChIP-Seq. These results provide new insight into fundamental mechanisms of gene traffic control, and point to an unexplored effect of anti-sense transcription on gene regulation via polymerase collision. Overall design: ChIP-Seq of RNA polymerase II was performed with WT and Elongain C deletion mutant (elc1∆) cells. 4H8 antibody against the Rpb1 C-terminal domain was used for RNA polymerase II immunoprecipitation, whilst mouse IgG antibody was used for control immunoprecipitations. Two biological replicates were performed for both WT and elc1∆.ENARNA Polymerase B, DNA-templated, cellular transcription, transcription, DNA-Dependent RNA Polymerase II, DNA-dependent., DNA Dependent RNA Polymerase II, transcription from bacterial-type RNA polymerase promoter, DNA-dependent transcription, RNA Pol II, bacterial transcriptionSaccharomyces oviformis, Baker's Yeasts, Yeast, S cerevisiae, Candida robusta, Saccharomyces capensis, Saccharomyces diastaticus, Saccharomyces cerevisiae 'var. diastaticus', Baker's, Brewer's, S. cerevisiae., baker's yeast, Mycoderma cerevisiae, Baker, Baker's Yeast, brewer's yeast, Saccharomyces uvarum var. melibiosus, Brewer's Yeast, Saccharomyces italicus, Baker Yeast0.00.00.00.00.00falseSaccharomyces cerevisiaeRNA polymerase II collision interrupts convergent transcription (ChIP-seq)2022-05-122013-05-31PRJNA167771GSE38382230412864932