[Isolation and characteristics of endonuclease tightly bound to alpha-polymerase from the rat liver].
Three major polypeptides are found in purified DNA polymerase alpha from rat liver: 160, 77 and 58 kDa. The electrophoretic analysis has identified polypeptide 160 kDa as the catalytically active subunit of DNA polymerase alpha. The other two polypeptides showed no DNA polymerase activity. Individual polypeptide p77 kDa purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to produce antibodies in rabbits. Immunoblot analysis indicated that the complex DNA polymerase alpha-3'-5'-exonuclease contained polypeptide p77 kDa. To elucidate the function of the p77 kDa protein we have prepared an immunoabsorbent column with antibodies against the p77 kDa polypeptide. The antibody column purified p77 kDa protein was homogeneous according to sodium dodecyl sulfate gel electrophoresis. The activity of alpha-polymerase was increased approximately 10-fold as a result of purification of DNA polymerase alpha from the p77 kDa protein. The in vitro experiments showed the identity of the p77 kDa polypeptide to endonuclease. It cleaved both single-stranded and double-stranded DNA. The function of endonuclease p77 kDA in complex with DNA polymerase alpha remains obscure.
Timchenko
N A
NA
Shevelev
I V
IV
Drobchenko
S N
SN
Zhuchenko
O P
OP
Krutiakov
V M
VM
D000818:Animals; D004257:DNA Polymerase II / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000302:isolation & purification* / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000032:analysis; D010455:Peptides / Q000032:analysis; D051381:Rats
D009097:Multienzyme Complexes; D010455:Peptides; D004257:DNA Polymerase II; D004720:Endonucleases
Vydelenie i kharakteristika éndonukleazy, zhestko sviazannoĭ s alpha-polimerazoĭ pecheni krysy.