GSE108673
RNA polymerase II-Independent Recruitment of SPT6L at Transcription Start Sites in Arabidopsis
SPT6 is a conserved elongation factor that is associated with phosphorylated RNA polymerase II (RNAPII) during transcription. Recent transcriptome analysis in yeast mutants revealed its potential role in the control of transcription initiation at genic promoters. However, the mechanism by which this is achieved and how this is linked to elongation remains to be elucidated. Here, we present the genome-wide occupancy of Arabidopsis SPT6-like (SPT6L) and demonstrate its conserved role in facilitating RNAPII occupancy across transcribed genes. We also further demonstrate that SPT6L enrichment is unexpectedly shifted, from gene body to transcription start site (TSS), when its association with RNAPII is disrupted. Protein domains, required for proper function and enrichment of SPT6L on chromatin, are subsequently identified. Finally, our results suggest that recruitment of SPT6L at TSS is indispensable for its spreading along the gene body during transcription. These findings provide new insights into the mechanisms underlying SPT6L recruitment in transcription and shed light on the coordination between transcription initiation and elongation. Overall design: Examination of RNA Polymerase II genome-wide binding in 10 days WT, spt6l, and spt6lSPT6L?SH2-GFP seedlings. After introducing ProSPT6L:SPT6L-GFP into spt6l seedlings, the genome-wide occupancy of SPT6L was examined. And then, a serial domain deletion of SPT6L was conducted and introduced into spt6l background. Finally, the genome-wide occupancy of SPT6L?HtH, SPT6L?YqgF, SPT6L?SH2, SPT6L?HtH?SH2, and SPT6L?YqgF?SH2 were performed.