共 30 条
Biochemical analysis of components of the pre-replication complex of Archaeoglobus fulgidus
被引:51
作者:

Grainge, I
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h-index: 0
机构:
Canc Res UK, Clare Hall Labs, London Res Inst, Potters Bar EN6 3LD, Herts, England Canc Res UK, Clare Hall Labs, London Res Inst, Potters Bar EN6 3LD, Herts, England

Scaife, S
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机构:
Canc Res UK, Clare Hall Labs, London Res Inst, Potters Bar EN6 3LD, Herts, England Canc Res UK, Clare Hall Labs, London Res Inst, Potters Bar EN6 3LD, Herts, England

Wigley, DB
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h-index: 0
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Canc Res UK, Clare Hall Labs, London Res Inst, Potters Bar EN6 3LD, Herts, England Canc Res UK, Clare Hall Labs, London Res Inst, Potters Bar EN6 3LD, Herts, England
机构:
[1] Canc Res UK, Clare Hall Labs, London Res Inst, Potters Bar EN6 3LD, Herts, England
关键词:
D O I:
10.1093/nar/gkg662
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
The eukaryotic pre-replication complex is assembled at replication origins in a reaction called licensing. Licensing involves the interactions of a variety of proteins including the origin recognition complex (ORC), Cdc6 and the Mcm2-7 helicase, homologues of which are also found in archaea. The euryarchaeote Archaeoglobus fulgidus encodes two genes with homology to Orc/Cdc6 and a single Mcm homologue. The A.fulgidus Mcm protein and one Orc/Cdc6 homologue have been purified and investigated in vitro. The Mcm protein is an ATP-dependent, hexameric helicase that can unwind between 200 and 400 bp of duplex DNA. Deletion of 112 amino acids from the N-terminus of A.f Mcm produced a protein, which was still capable of forming a hexamer, was competent in DNA binding and was able to unwind at least 1 kb of duplex DNA. The purified Orc/Cdc6 homologue was also able to bind DNA. Both Mcm and Orc/Cdc6 show a preference for specific DNA structures, namely molecules containing a single stranded bubble that mimics early replication intermediates. Nuclease protection showed that the binding sites for Mcm and Orc/Cdc6 overlap. The Orc/Cdc6 protein bound more tightly to these substrates and was able to displace pre-bound Mcm hexamer.
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页码:4888 / 4898
页数:11
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