Basu, A., Dong, B., Krainer, A. R., Howe, C. C. (February 1997) The intracisternal A-particle proximal enhancer-binding protein activates transcription and is identical to the RNA- and DNA-binding protein p54nrb/NonO. Molecular and Cellular Biology, 17 (2). pp. 677-86. ISSN 0270-7306 (Print)0270-7306
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Abstract
The long terminal repeats of murine intracisternal A particles (IAPs) contain an IAP proximal enhancer (IPE) element that is inactive in murine F9 embryonal carcinoma cells and active in the parietal endoderm cell line PYS-2. The element binds efficiently to a 60-kDa IPE-binding protein (IPEB) present in PYS-2 cells but poorly to F9 proteins, suggesting a role for IPEB in regulating IAP expression. We have purified calf thymus IPEB, which binds to the IPE and transactivates a reporter gene in HeLa cell extracts. Based on the peptide sequence of the purified calf IPEB, we have cloned a 420-bp cDNA and showed that the encoded protein is the homolog of human p54nrb and mouse NonO, which are characterized by the presence of two RNA recognition motifs. We show that p54nrb is an IPE-binding transcription activator with its DNA-binding and activation domains in the N- and C-terminal halves, respectively. The activation domain of p54nrb is active in HeLa, PYS-2, and F9 cells, whereas p54nrb as a whole molecule is active in HeLa and PYS-2 cells but not in F9 cells. Thus, the lack of activity of p54nrb in F9 cells is due to an ineffective DNA-binding domain. We demonstrate that p54nrb also binds to a pre-mRNA. Based on the close sequence relatedness of this protein to PSF, which is required for pre-mRNA splicing in vitro, we discuss the possibility that p54nrb has dual roles in transcription and splicing.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Animals Cattle Cell Line Cloning, Molecular Cyclin D1 DNA-Binding Proteins/chemistry/*genetics/isolation & purification/metabolism Enhancer Elements, Genetic/genetics Genes, Intracisternal A-Particle/*genetics HeLa Cells Humans Mice Molecular Weight *Nuclear Matrix-Associated Proteins Nuclear Proteins/*genetics Octamer Transcription Factors Proto-Oncogene Proteins/genetics RNA Precursors/metabolism RNA-Binding Proteins/chemistry/*genetics/isolation & purification Recombinant Fusion Proteins Sequence Homology, Amino Acid Thymus Gland Trans-Activators/chemistry/*genetics/isolation & purification/metabolism Transfection |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > transcription bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > RNA binding protein |
| CSHL Authors: | |
| Communities: | CSHL labs > Krainer lab |
| Depositing User: | Matt Covey |
| Date: | February 1997 |
| Date Deposited: | 11 Mar 2014 19:53 |
| Last Modified: | 11 Mar 2014 19:53 |
| PMCID: | PMC231793 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29594 |
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