Splicing and transcription-associated proteins PSF and p54(nrb)/NonO bind to the RNA polymerase II CTD

Emili, A., Shales, M., McCracken, S., Xie, W. J., Tucker, P. W., Kobayashi, R., Blencowe, B. J., Ingles, C. J. (September 2002) Splicing and transcription-associated proteins PSF and p54(nrb)/NonO bind to the RNA polymerase II CTD. RNA, 8 (9). pp. 1102-1111. ISSN 1355-8382

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URL: http://www.ncbi.nlm.nih.gov/pubmed/12358429

Abstract

The carboxyl-terminal domain (CTD) of the largest subunit of eukaryotic RNA polymerase II (pol II) plays an important role in promoting steps of pre-mRNA processing. To identify proteins in human cells that bind to the CTD and that could mediate its functions in pre-mRNA processing, we used the mouse CTD expressed in bacterial cells in affinity chromatography experiments. Two proteins present in HeLa cell extract, the splicing and transcription-associated factors, PSF and p54nrb/NonO, bound specifically and could be purified to virtual homogeneity by chromatography on immobilized CTD matrices. Both hypo- and hyperphosphorylated CTD matrices bound these proteins with similar selectivity. PSF and p54nrb/NonO also copurified with a holoenzyme form of pol II containing hypophosphorylated CTD and could be coimmunoprecipitated with antibodies specific for this and the hyperphosphorylated form of pol II. That PSF and p54nrb/NonO promoted the binding of RNA to immobilized CTD matrices suggested these proteins can interact with the CTD and RNA simultaneously. PSF and p54nrb/NonO may therefore provide a direct physical link between the pol II CTD and pre-mRNA processing components, at both the initiation and elongation phases of transcription.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > RNA polymerase
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > splicing factor
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > transcription factor
CSHL Authors:
Communities: CSHL labs > Kobayashi lab
Depositing User: Matt Covey
Date: September 2002
Date Deposited: 08 Jan 2014 20:50
Last Modified: 08 Jan 2014 20:50
PMCID: PMC1370324
Related URLs:
URI: http://repository.cshl.edu/id/eprint/28701

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