Depletion of REF/Aly alters gene expression and reduces RNA polymerase II occupancy

Sarah H. Stubbs, Nicholas K. Conrad

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Pre-mRNA processing is mechanistically linked to transcription with RNA pol II serving as a platform to recruit RNA processing factors to nascent transcripts. The TREX complex member, REF/Aly, has been suggested to play roles in transcription and nuclear RNA stability in addition to its more broadly characterized role in mRNA export. We employed RNA-seq to identify a subset of transcripts with decreased expression in both nuclear and cytoplasmic fractions upon REF/Aly knockdown, which implies that REF/Aly affects their expression upstream of its role in mRNA export. Transcription inhibition experiments and metabolic labeling assays argue that REF/Aly does not affect stability of selected candidate transcripts. Instead, ChIP assays and nuclear run-on analysis reveal that REF/Aly depletion diminishes the transcription of these candidate genes. Furthermore, we determined that REF/Aly binds directly to candidate transcripts, supporting a direct effect of REF/Aly on candidate gene transcription. Taken together, our data suggest that the importance of REF/Aly is not limited to RNA export, but that REF/Aly is also critical for gene expression at the level of transcription. Our data are consistent with the model that REF/Aly is involved in linking splicing with transcription efficiency.

Original languageEnglish (US)
Pages (from-to)504-519
Number of pages16
JournalNucleic Acids Research
Volume43
Issue number1
DOIs
StatePublished - Sep 1 2015

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RNA Polymerase II
RNA
Gene Expression
Nuclear RNA
Messenger RNA
RNA Precursors
RNA Stability
Genes

ASJC Scopus subject areas

  • Genetics

Cite this

Depletion of REF/Aly alters gene expression and reduces RNA polymerase II occupancy. / Stubbs, Sarah H.; Conrad, Nicholas K.

In: Nucleic Acids Research, Vol. 43, No. 1, 01.09.2015, p. 504-519.

Research output: Contribution to journalArticle

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