A constitutive heat shock element-binding factor is immunologically identical to the Ku autoantigen Journal Article
| Authors: | Kim, D.; Ouyang, H.; Yang, S. H.; Nussenzweig, A.; Burgman, P.; Li, G. C. |
| Article Title: | A constitutive heat shock element-binding factor is immunologically identical to the Ku autoantigen |
| Abstract: | Analysis of the heat shock element (HSE)-binding proteins in extracts of rodent cells, during heat shock and their post-heat shock recovery, indicates that the regulation of heat shock response involves a constitutive HSE- binding factor (CHBF), in addition to the heat-inducible heat shock factor HSF1. We purified the CHBF to apparent homogeneity from HeLa cells using column chromatographic techniques including an HSE oligonucleotide affinity column. The purified CHBF consists of two polypeptides with apparent molecular masses of 70 and 86 kDa. Immunoblot and gel mobility shift analysis verify that CHBF is identical or closely related to the Ku autoantigen. The DNA binding characteristics of CHBF to double-stranded or single-stranded DNA are similar to that of Ku autoantigen. In gel mobility shift analysis using purified CHBF and recombinant human HSF1, CHBF competes with HSF1 for the binding of DNA sequences containing HSEs in vitro. Furthermore, when Rat-1 cells were co-transfected with human Ku expression vectors and the hsp70- promoter-driven luciferase reporter gone, thermal induction of luciferase is significantly suppressed relative to cells transfected with only the hsp70- luciferase construct. These data suggest a role of CHBF (or Ku protein) in the regulation of heat response in vivo. |
| Keywords: | human cell; promoter region; dna-binding proteins; comparative study; animal; cell line; luciferase; hela cell; hela cells; transfection; nuclear proteins; gene expression regulation; antibodies, monoclonal; dna; molecular sequence data; recombinant proteins; substrate specificity; immunoblotting; reporter gene; autoantigen; base sequence; binding sites; rats; dna, single-stranded; ultraviolet rays; molecular weight; protein dna interaction; thermoregulation; oligodeoxyribonucleotides; binding, competitive; chromatography, affinity; heat shock protein; heat; human; priority journal; article; support, u.s. gov't, p.h.s.; heat stroke |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 270 |
| Issue: | 25 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1995-06-23 |
| Start Page: | 15277 |
| End Page: | 15284 |
| Language: | English |
| DOI: | 10.1074/jbc.270.25.15277 |
| PUBMED: | 7797514 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 28 August 2018 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work