The human homologue of fission Yeast cdc27, p66, is a component of active human DNA polymerase delta.
J Biochem
; 129(5): 699-708, 2001 May.
Article
en En
| MEDLINE
| ID: mdl-11328591
ABSTRACT
An essential eukaryotic DNA polymerase, DNA polymerase delta (pol delta), synthesizes DNA processively in the presence of proliferating cell nuclear antigen (PCNA). Recently, a 66 kDa polypeptide (p66) that displays significant homology within its PCNA binding domain to that of fission yeast cdc27 was identified as a component of mouse and calf thymus pol delta. Our studies show that p66 interacts tightly with other subunits of pol delta during size fractionation of human cell extracts, and co-immunoprecipitates with these subunits along with PCNA-dependent polymerase activity. Active human pol delta could be reconstituted by co-expressing p125, p50, and p66 recombinant baculoviruses, but not by co-expressing p125 and p50 alone. Interaction studies demonstrated that p66 stabilizes the association between p125 and p50. Pull-down assays with PCNA-linked beads demonstrated that p66 increases the overall affinity of pol delta for PCNA. These results indicate that p66 is a functionally important subunit of human pol delta that stabilizes the pol delta complex and increases the affinity of pol delta for PCNA.
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Banco de datos:
MEDLINE
Asunto principal:
Péptidos
/
Antígeno Nuclear de Célula en Proliferación
/
Proteínas de Ciclo Celular
/
ADN Polimerasa III
Límite:
Animals
/
Humans
Idioma:
En
Año:
2001
Tipo del documento:
Article