Purification and characterization of p27, a protein from hepatocyte chromatin. Evidence suggesting that it binds selectively to guanine-rich single-stranded DNA.
A protein designated p27 that binds preferentially to single-stranded DNA rich in guanine tracts was purified to near homogeneity from rabbit hepatocyte non-histone protein extract. Purified p27 migrated as a 27 kDa polypeptide on denaturing SDS-PAGE and displayed a native molecular mass of approximately 155 kDa on Sephadex G-150 or Sepharose 6B-Cl gel filtration columns. Gel shift analysis indicated that maximum binding of p27 to single-stranded DNA required the presence of tracts of four or more contiguous guanine residues. The lowest found dissociation constant, 1.4 x 10(-8) M/l, was for single-stranded DNA that contained a (dG)17 run.
Rahat
M A
MA
Unit of Biochemistry, Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Bat Galim, Haifa.
Fry
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M
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002843:Chromatin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004247:DNA; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000737:chemistry* / Q000166:cytology; D008969:Molecular Sequence Data; D011817:Rabbits
D002843:Chromatin; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D004247:DNA