title: Isolation and characterization of a DNA helicase from cytosolic extracts of calf thymus.
abstract: A DNA helicase has been isolated from calf thymus tissue. The enzyme was enriched from crude cytosolic extracts by batchwise chromatography on phosphocellulose, followed by 35% ammonium sulfate precipitation, and subsequent chromatography on phenyl-Sepharose, single-stranded DNA cellulose, and AcA 44 gel filtration. The DNA helicase had a Stokes' radius of about 45 A and a sedimentation coefficient of 4.3 S. The most purified fractions contained three polypeptides with apparent molecular weights of 110, 65, and 34 kDa. UV crosslinking with radioactive dATP stained all three major polypeptides. The helicase catalyzed the unwinding of a DNA primer from a single-stranded DNA template in an ATP- or dATP-dependent manner. DNA unwinding was also observed with CTP or dCTP, but with reduced efficiency. The helicase translocated from 3' to 5' on the single-stranded template it was bound to. Relationships between this DNA helicase and other calf thymus helicases will be discussed.
authors.affiliation: German Primate Center, Department of Virology and Immunology, Göttingen.
authors.forename: S
authors.initials: S
authors.lastname: Zhang
authors.forename: F
authors.initials: F
authors.lastname: Grosse
mesh_terms: D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003570:Cytidine Triphosphate / Q000502:physiology; D003600:Cytosol / Q000201:enzymology*; D004247:DNA / Q000378:metabolism; D004265:DNA Helicases / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D013950:Thymus Gland / Q000201:enzymology
chemical_list: D003570:Cytidine Triphosphate; D000255:Adenosine Triphosphate; D004247:DNA; D004265:DNA Helicases