Spt5 Regulates Sense and Antisense Transcription Genome-Wide

Abstract

Spt5 is the only transcription elongation factor conserved in all three domains of life, but its molecular mechanisms are not yet thoroughly studied genomically. From an inducible depletion strain, we sequenced nascent transcripts (NET-seq), mature mRNA (RNA-seq), and RNA polymerase II-associated chromatin (ChIP-seq) to elucidate general effects of Spt5 on transcription. These show an increase in 5’ CDS antisense transcription by RNA-seq and NET-seq and a general accumulation of RNA Pol II at the 5’ ends of genes by exogenous spike-in-normalized ChIP-seq. That similar antisense transcripts appear in RNA-seq and NET-seq indicates that novel antisense transcripts are resulting from new transcription rather than aberrant decay. These results provide insight into how Spt5 functions to facilitate RNA Pol II elongation in diverse organisms.