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-Structure paper
タイトル | DNA induces conformational changes in a recombinant human minichromosome maintenance complex. |
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ジャーナル・号・ページ | J Biol Chem, Vol. 290, Issue 12, Page 7973-7979, Year 2015 |
掲載日 | 2015年3月20日 |
著者 | Emma L Hesketh / Richard P Parker-Manuel / Yuriy Chaban / Rabab Satti / Dawn Coverley / Elena V Orlova / James P J Chong / |
PubMed 要旨 | ATP-dependent DNA unwinding activity has been demonstrated for recombinant archaeal homohexameric minichromosome maintenance (MCM) complexes and their yeast heterohexameric counterparts, but in ...ATP-dependent DNA unwinding activity has been demonstrated for recombinant archaeal homohexameric minichromosome maintenance (MCM) complexes and their yeast heterohexameric counterparts, but in higher eukaryotes such as Drosophila, MCM-associated DNA helicase activity has been observed only in the context of a co-purified Cdc45-MCM-GINS complex. Here, we describe the production of the recombinant human MCM (hMCM) complex in Escherichia coli. This protein displays ATP hydrolysis activity and is capable of unwinding duplex DNA. Using single-particle asymmetric EM reconstruction, we demonstrate that recombinant hMCM forms a hexamer that undergoes a conformational change when bound to DNA. Recombinant hMCM produced without post-translational modifications is functional in vitro and provides an important tool for biochemical reconstitution of the human replicative helicase. |
リンク | J Biol Chem / PubMed:25648893 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 23.0 Å |
構造データ | EMDB-2872: EMDB-2873: |
由来 |
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