Bennett, C. F., Spector, D. L., Yeoman, L. C. (February 1986) Nonhistone protein BA is a glutathione S-transferase localized to interchromatinic regions of the cell nucleus. Journal of Cell Biology, 102 (2). pp. 600-9. ISSN 0021-9525
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Abstract
A DNA-binding nonhistone protein, protein BA, was previously demonstrated to co-localize with U-snRNPs within discrete nuclear domains (Bennett, F. C., and L. C. Yeoman, 1985, Exp. Cell Res., 157:379-386). To further define the association of protein BA and U-snRNPs within these discrete nuclear domains, cells were fractionated in situ and the localization of the antigens determined by double-labeled immunofluorescence. Protein BA was extracted from the nucleus with the 2.0 M NaCl soluble chromatin fraction, while U-snRNPs were only partially extracted from the 2.0 M NaCl-resistant nuclear structures. U-snRNPs were extracted from the residual nuclear material by combined DNase I/RNase A digestions. Using an indirect immunoperoxidase technique and electron microscopy, protein BA was localized to interchromatinic regions of the cell nucleus. Protein BA was noted to share a number of chemical and physical properties with a family of cytoplasmic enzymes, the glutathione S-transferases. Comparison of the published amino acid composition of protein BA and glutathione S-transferases showed marked similarities. Nonhistone protein BA isolated from saline-EDTA nuclear extracts exhibited glutathione S-transferase activity with a variety of substrates. Substrate specificity and subunit analysis by SDS polyacrylamide gel electrophoresis revealed that it was a mixture of several glutathione S-transferase isoenzymes. Protein BA isolated from rat liver chromatin was shown by immunoblotting and peptide mapping techniques to be two glutathione S-transferase isoenzymes composed of the Yb and Yb' subunits. Glutathione S-transferase Yb subunits were demonstrated to be both nuclear and cytoplasmic proteins by indirect immunolocalization on rat liver cryosections. The identification of protein BA as glutathione S-transferase suggests that this family of multifunctional enzymes may play an important role in those nuclear domains containing U-snRNPs.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Amino Acids analysis Animals Cell Compartmentation Cell Nucleus enzymology ultrastructure Chromosomal Proteins Non-Histone metabolism Electrophoresis Polyacrylamide Gel Fluorescent Antibody Technique Glutathione Transferase analysis metabolism Immunoenzyme Techniques Macromolecular Substances Microscopy Electron Molecular Weight Peptide Fragments analysis Rats Ribonucleoproteins metabolism Ribonucleoproteins Small Nuclear |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein |
| CSHL Authors: | |
| Communities: | CSHL labs > Spector lab |
| Depositing User: | Brian Soldo |
| Date: | February 1986 |
| Date Deposited: | 04 Apr 2012 20:14 |
| Last Modified: | 29 Jan 2015 20:51 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/25643 |
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