Nascent RNA sequencing reveals widespread pausing and divergent initiation at human promoters

Science. 2008 Dec 19;322(5909):1845-8. doi: 10.1126/science.1162228. Epub 2008 Dec 4.

Abstract

RNA polymerases are highly regulated molecular machines. We present a method (global run-on sequencing, GRO-seq) that maps the position, amount, and orientation of transcriptionally engaged RNA polymerases genome-wide. In this method, nuclear run-on RNA molecules are subjected to large-scale parallel sequencing and mapped to the genome. We show that peaks of promoter-proximal polymerase reside on approximately 30% of human genes, transcription extends beyond pre-messenger RNA 3' cleavage, and antisense transcription is prevalent. Additionally, most promoters have an engaged polymerase upstream and in an orientation opposite to the annotated gene. This divergent polymerase is associated with active genes but does not elongate effectively beyond the promoter. These results imply that the interplay between polymerases and regulators over broad promoter regions dictates the orientation and efficiency of productive transcription.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Line
  • CpG Islands
  • DNA-Directed RNA Polymerases / metabolism*
  • Genome, Human
  • Humans
  • Nucleosomes / metabolism
  • Promoter Regions, Genetic*
  • RNA Polymerase II / metabolism*
  • RNA, Antisense / genetics
  • RNA, Antisense / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sequence Analysis, RNA*
  • Transcription Initiation Site
  • Transcription, Genetic*

Substances

  • Nucleosomes
  • RNA, Antisense
  • RNA, Messenger
  • RNA Polymerase II
  • DNA-Directed RNA Polymerases

Associated data

  • GEO/GSE13518