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Purification of a novel factor which binds to the mouse α2 (I) collagen promoter
Author(s) -
Hatamochi Atsushi,
de Crombrugghe Benoit,
Krieg Thomas
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81014-q
Subject(s) - microbiology and biotechnology , dna , affinity chromatography , transcription factor , dna binding protein , agarose , polyacrylamide gel electrophoresis , electrophoretic mobility shift assay , gel electrophoresis , 3t3 cells , sepharose , biology , nuclear protein , biochemistry , chemistry , gene , enzyme , transfection
We have identified and purified a DNA binding protein which specifically binds to a segment of the mouse α2 (I) collagen promoter between −420 and −399 bp upstream of the start of transcription. Purification included heparin‐agarose and sequence‐specific DNA‐affinity chromatography, followed by SDS‐PAGE and renaturation of the DNA binding activity after elution from SDS‐polyacrylamide gel. The DNA binding activity resides in two species of 42 kDa and 40 kDa, respectively. The levels of DNA binding activity of this factor, which has been tentatively designated as ColFl, are considerably higher in nuclear extracts of NIH‐3T3 fibroblasts than in nuclear extracts from epidermal cells, lymphoid cells and transformed NIH‐3T3 fibroblasts.