We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
A ring-opening mechanism for DNA binding in the central channel of the T7 helicase-primase protein.
- Authors
Ahnert, Peter; Picha, Kristen Moore; Patel, Smita S.
- Abstract
We have investigated the mechanism of binding single-stranded DNA (ssDNA) into the central channel of the ring-shaped T7 gp4A′ helicaseprimase hexamer. Presteadystate kinetic studies show a facilitated five-step mechanism and provide understanding of how a ring-shaped helicase can be loaded on the DNA during the initiation of replication. The effect of a primase recognition sequence on the observed kinetics suggests that binding to the helicase DNA-binding site is facilitated by transient binding to the primase DNA-binding site, which is proposed to be a loading site. The proposed model involves the fast initial binding of the DNA to the primase site on the outside of the helicase ring, a fast conformational change, a ring-opening step, migration of the DNA into the central channel of the helicase ring, and ring closure. Although an intermediate protein-DNA complex is kinetically stable, only the last species in the five-step mechanism is poised to function as a helicase at the unwinding junction.
- Subjects
DNA-protein interactions; DNA-ligand interactions; PROTEIN binding; DNA helicases; DNA; GENETIC research
- Publication
EMBO Journal, 2000, Vol 19, Issue 13, p3418
- ISSN
0261-4189
- Publication type
Article
- DOI
10.1093/emboj/19.13.3418