DSIF and NELF interact with Integrator to specify the correct post-transcriptional fate of snRNA genes

Junichi Yamamoto; Yuri Hagiwara; Kunitoshi Chiba; Tomoyasu Isobe; Takashi Narita; Hiroshi Handa; Yuki Yamaguchi

doi:10.1038/ncomms5263

DSIF and NELF interact with Integrator to specify the correct post-transcriptional fate of snRNA genes

Nat Commun. 2014 Jun 27:5:4263. doi: 10.1038/ncomms5263.

Authors

Junichi Yamamoto¹, Yuri Hagiwara¹, Kunitoshi Chiba¹, Tomoyasu Isobe¹, Takashi Narita², Hiroshi Handa³, Yuki Yamaguchi⁴

Affiliations

¹ Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226-8501, Japan.
² Graduate School of Frontier Biosciences, Osaka University, Suita 565-0871, Japan.
³ Department of Nanoparticle Translational Research, Tokyo Medical University, Tokyo 160-8402, Japan.
⁴ 1] Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226-8501, Japan [2] PRESTO, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan.

PMID: 24968874
DOI: 10.1038/ncomms5263

Abstract

The elongation factors DSIF and NELF are responsible for promoter-proximal RNA polymerase II (Pol II) pausing. NELF is also involved in 3' processing of replication-dependent histone genes, which produce non-polyadenylated mRNAs. Here we show that DSIF and NELF contribute to the synthesis of small nuclear RNAs (snRNAs) through their association with Integrator, the large multisubunit complex responsible for 3' processing of pre-snRNAs. In HeLa cells, Pol II, Integrator, DSIF and NELF accumulate at the 3' end of the U1 snRNA gene. Knockdown of NELF results in misprocessing of U1, U2, U4 and U5 snRNAs, while DSIF is required for proper transcription of these genes. Knocking down NELF also disrupts transcription termination and induces the production of polyadenylated U1 transcripts caused by an enhanced recruitment of cleavage stimulation factor. Our results indicate that NELF plays a key role in determining the post-transcriptional fate of Pol II-transcribed genes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carrier Proteins / metabolism
DNA-Binding Proteins / metabolism
Endoribonucleases / metabolism
Gene Knockdown Techniques
HeLa Cells
Humans
Intracellular Signaling Peptides and Proteins
Nuclear Proteins / genetics
Nuclear Proteins / metabolism*
RNA Polymerase II / metabolism*
RNA, Small Nuclear / metabolism*
RNA-Binding Proteins
Ribosomal Proteins / metabolism
Transcription Factors / genetics
Transcription Factors / metabolism*
Transcriptional Elongation Factors / genetics
Transcriptional Elongation Factors / metabolism*
Tumor Suppressor Proteins / metabolism

Substances

Carrier Proteins
DNA-Binding Proteins
INTS12 protein, human
INTS3 protein, human
INTS4 protein, human
INTS6 protein, human
Intracellular Signaling Peptides and Proteins
NSMF protein, human
Nuclear Proteins
RNA, Small Nuclear
RNA-Binding Proteins
Ribosomal Proteins
SUPT5H protein, human
Transcription Factors
Transcriptional Elongation Factors
Tumor Suppressor Proteins
U1 small nuclear RNA
U2 small nuclear RNA
U4 small nuclear RNA
U5 small nuclear RNA
RNA Polymerase II
Endoribonucleases
IntS9 protein, human