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Pathway of Complex Formation between DNA and Three Subunits of CBF/NF-Y
- Source :
- Journal of Biological Chemistry. 273:31590-31598
- Publication Year :
- 1998
- Publisher :
- Elsevier BV, 1998.
-
Abstract
- In this study, we used a photocross-linking method to identify specific contact of CCAAT-binding factor (CBF) subunits in a CBF-DNA complex. The analysis showed that all three subunits in the CBF-DNA complex were cross-linked to DNA and that CBF-B and CBF-C were cross-linked more strongly than CBF-A. None of the CBF-A and CBF-C subunits, which together formed a CBF-A/CBF-C heterodimer, were cross-linked without CBF-B; in contrast, CBF-B was cross-linked in the absence of CBF-A/CBF-C. No subunit of heterotrimeric CBF containing DNA-binding domain mutant of either CBF-B or CBF-C was cross-linked to DNA, and interestingly, cross-linking of CBF-B that occurred without CBF-A/CBF-C was inhibited in presence of mutant CBF-C/CBF-A heterodimer. Altogether, these results indicated that the specific DNA contact surface of each CBF subunit is generated as a result of interaction between CBF-B and CBF-A/CBF-C heterodimer and that the three CBF subunits interact interdependently with DNA to form a CBF-DNA complex. Equilibrium interactions among the three CBF subunits and between CBF subunits and DNA were studied by electrophoretic mobility shift assay. This showed that at equilibrium DNA-binding conditions, the CBF-A/CBF-C heterodimer is very stable, but association between CBF-B and CBF-A/CBF-C is very weak. The nature of the association of CBF-B with CBF-A/CBF-C was also revealed by studying the inhibition of CBF-DNA complex formation by the mutant CBF-B. This study indicated that the association between CBF-B and CBF-A/CBF-C is stabilized upon interaction with DNA, a process likely to favor formation of a high-affinity CBF-DNA complex.
- Subjects :
- Mutation
Protein subunit
Mutant
Cell Biology
Biology
medicine.disease_cause
Biochemistry
Molecular biology
chemistry.chemical_compound
Protein structure
nervous system
chemistry
Heterotrimeric G protein
cardiovascular system
medicine
Biophysics
Protein–DNA interaction
Electrophoretic mobility shift assay
Molecular Biology
DNA
circulatory and respiratory physiology
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 273
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi...........272e8240c487a0cd76c4c9d5ebeffa8b