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-Structure paper
タイトル | Architecture and ssDNA interaction of the Timeless-Tipin-RPA complex. |
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ジャーナル・号・ページ | Nucleic Acids Res, Vol. 42, Issue 20, Page 12912-12927, Year 2014 |
掲載日 | 2014年11月10日 |
著者 | Justine Witosch / Eva Wolf / Naoko Mizuno / |
PubMed 要旨 | The Timeless-Tipin (Tim-Tipin) complex, also referred to as the fork protection complex, is involved in coordination of DNA replication. Tim-Tipin is suggested to be recruited to replication forks ...The Timeless-Tipin (Tim-Tipin) complex, also referred to as the fork protection complex, is involved in coordination of DNA replication. Tim-Tipin is suggested to be recruited to replication forks via Replication Protein A (RPA) but details of the interaction are unknown. Here, using cryo-EM and biochemical methods, we characterized complex formation of Tim-Tipin, RPA and single-stranded DNA (ssDNA). Tim-Tipin and RPA form a 258 kDa complex with a 1:1:1 stoichiometry. The cryo-EM 3D reconstruction revealed a globular architecture of the Tim-Tipin-RPA complex with a ring-like and a U-shaped domain covered by a RPA lid. Interestingly, RPA in the complex adopts a horse shoe-like shape resembling its conformation in the presence of long ssDNA (>30 nucleotides). Furthermore, the recruitment of the Tim-Tipin-RPA complex to ssDNA is modulated by the RPA conformation and requires RPA to be in the more compact 30 nt ssDNA binding mode. The dynamic formation and disruption of the Tim-Tipin-RPA-ssDNA complex implicates the RPA-based recruitment of Tim-Tipin to the replication fork. |
リンク | Nucleic Acids Res / PubMed:25348395 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 17.0 Å |
構造データ | EMDB-2789: |
由来 |
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