p54nrb is a component of the snRNP-free U1A (SF-A) complex that promotes pre-mRNA cleavage during polyadenylation

Songchun Liang, Carol Lutz

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The U1 snRNP-A (U1A) protein has been known for many years as a component of the U1 snRNP. We have previously described a form of U1A present in human cells in significant amounts that is not associated with the U1 snRNP or U1 RNA but instead is part of a novel complex of non-snRNP proteins that we have termed snRNP-free U1A, or SF-A. Antibodies that specifically recognize this complex inhibit in vitro splicing and polyadenylation of pre-mRNA, suggesting that this complex may play an important functional role in these mRNA-processing activities. This finding was underscored by the determination that one of the components of this complex is the polypyrimidine-tract-binding protein-associated splicing factor, PSF. In order to further our studies on this complex and to determine the rest of the components of the SF-A complex, we prepared several stable HeLa cell lines that overexpress a tandem-affinity- purification-tagged version of U1A (TAP-tagged U1A). Nuclear extract was prepared from one of these cell lines, line 107, and affinity purification was performed along with RNase treatment. We have used mass spectrometry analysis to identify the candidate factors that associate with U1A. We have now identified and characterized PSF, p54nrb, and p68 as novel components of the SF-A complex. We have explored the function of this complex in RNA processing, specifically cleavage and polyadenylation, by performing immunodepletions followed by reconstitution experiments, and have found that p54nrb is critical. Published by Cold Spring Harbor Laboratory Press.

Original languageEnglish (US)
Pages (from-to)111-121
Number of pages11
JournalRNA
Volume12
Issue number1
DOIs
StatePublished - Jan 1 2006

Fingerprint

U1 Small Nuclear Ribonucleoproteins
Small Nuclear Ribonucleoproteins
Polyadenylation
RNA Precursors
Cell Line
Ribonucleases
HeLa Cells
Mass Spectrometry
Proteins
RNA
Messenger RNA
Antibodies

All Science Journal Classification (ASJC) codes

  • Molecular Biology

Keywords

  • Polyadenylation
  • SF-A
  • U1 snRNP
  • U1A

Cite this

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p54nrb is a component of the snRNP-free U1A (SF-A) complex that promotes pre-mRNA cleavage during polyadenylation. / Liang, Songchun; Lutz, Carol.

In: RNA, Vol. 12, No. 1, 01.01.2006, p. 111-121.

Research output: Contribution to journalArticle

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