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Human p32, interacts with B subunit of the CCAAT-binding factor, CBF/NF-Y, and inhibits CBF-mediated transcription activation in vitro
- Source :
- Nucleic Acids Research. 32:3632-3641
- Publication Year :
- 2004
- Publisher :
- Oxford University Press (OUP), 2004.
-
Abstract
- To understand the role of the CCAAT-binding factor, CBF, in transcription, we developed a strategy to purify the heterotrimeric CBF complex from HeLa cell extracts using two successive immunoaffinity chromatography steps. Here we show that the p32 protein, previously identified as the ASF/SF2 splicing factor-associated protein, copurified with the CBF complex. Studies of protein–protein interaction demonstrated that p32 interacts specifically with CBF–B subunit and also associates with CBF–DNA complex. Cellular localization by immunofluorescence staining revealed that p32 is present in the cell throughout the cytosol and nucleus, whereas CBF is present primarily in the nucleus. A portion of the p32 colocalizes with CBF-B in the nucleus. Interestingly, reconstitution of p32 in an in vitro transcription reaction demonstrated that p32 specifically inhibits CBF-mediated transcription activation. Altogether, our study identified p32 as a novel and specific corepressor of CBF-mediated transcription activation in vitro.
- Subjects :
- Transcriptional Activation
Macromolecular Substances
Protein subunit
Biology
Chromatography, Affinity
Mitochondrial Proteins
Transcription (biology)
Heterotrimeric G protein
Genetics
medicine
Humans
Cellular localization
Cell Nucleus
Membrane Glycoproteins
Articles
Molecular biology
Recombinant Proteins
Receptors, Complement
Repressor Proteins
body regions
Protein Subunits
Cytosol
Cell nucleus
Hyaluronan Receptors
medicine.anatomical_structure
CCAAT-Binding Factor
nervous system
cardiovascular system
Carrier Proteins
Corepressor
Nucleus
HeLa Cells
circulatory and respiratory physiology
Subjects
Details
- ISSN :
- 13624962
- Volume :
- 32
- Database :
- OpenAIRE
- Journal :
- Nucleic Acids Research
- Accession number :
- edsair.doi.dedup.....e277963968e3ca15be7c5175b5eb892e