In:
Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 93, No. 23 ( 1996-11-12), p. 12896-12901
Abstract:
Human replication factor C (RFC, also called activator 1) is a
five-subunit protein complex (p140, p40, p38, p37, and p36) required for proliferating cell nuclear antigen (PCNA)-dependent processive DNA
synthesis catalyzed by DNA polymerase δ or ɛ. Here we report the reconstitution of the RFC complex from its five subunits simultaneously
overexpressed in baculovirus-infected insect cells. The purified baculovirus-produced RFC appears to contain equimolar levels of each
subunit and was shown to be functionally identical to its native counterpart in ( i ) supporting DNA polymerase
δ-catalyzed PCNA-dependent DNA chain elongation; ( ii )
catalyzing DNA-dependent ATP hydrolysis that was stimulated by PCNA and human single-stranded DNA binding protein; ( iii ) binding
preferentially to DNA primer ends; and ( iv )
catalytically loading PCNA onto singly nicked circular DNA and catalytically removing PCNA from these DNA molecules.
Type of Medium:
Online Resource
ISSN:
0027-8424
,
1091-6490
DOI:
10.1073/pnas.93.23.12896
Language:
English
Publisher:
Proceedings of the National Academy of Sciences
Publication Date:
1996
detail.hit.zdb_id:
209104-5
detail.hit.zdb_id:
1461794-8
SSG:
11
SSG:
12
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