A CCAAT DNA binding factor consisting of two different components that are both required for DNA binding.
A CCAAT-binding activity present in nuclear extracts of rat liver and NIH 3T3 fibroblasts was purified using, as assay, DNA binding to a segment of the mouse alpha 2(I) collagen promoter. The activity consists of two components, designated factors A and B, which are separated by ion exchange chromatography on either Mono Q or Mono S columns. Factor A is heat-sensitive, whereas factor B is heat-resistant. Both factors are required for DNA binding and both are present in the DNA protein complex. The A + B complex was extensively purified by heparin-agarose and sequence-specific affinity chromatography. The Mr of factor A is 39,000, whereas the Mr of factor B is 41,000 as determined by renaturation of a highly purified preparation after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Competition experiments indicate that this CCAAT-binding complex has a DNA sequence specificity that is different from those of other CCAAT-binding proteins.
Hatamochi
A
A
Laboratory of Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892.
Golumbek
P T
PT
Van Schaftingen
E
E
de Crombrugghe
B
B
D000818:Animals; D001483:Base Sequence; D002413:Cations, Divalent; D002467:Cell Nucleus / Q000032:analysis; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003094:Collagen / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005090:Exodeoxyribonucleases; D005347:Fibroblasts / Q000032:analysis*; D006358:Hot Temperature; D008099:Liver / Q000032:analysis*; D008970:Molecular Weight; D009841:Oligonucleotides / Q000302:isolation & purification* / Q000378:metabolism; D010447:Peptide Hydrolases / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D051381:Rats; D015032:Zinc / Q000494:pharmacology
D002413:Cations, Divalent; D009841:Oligonucleotides; D003094:Collagen; D004247:DNA; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III; D010447:Peptide Hydrolases; D015032:Zinc