Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA–RuvB protein complex | Zendy

Yong-Woon Han | Zendy; Tomomi Tani | Zendy; Masahito Hayashi | Zendy; Takashi Hishida | Zendy; Hiroshi Iwasaki | Zendy; Hideo Shinagawa | Zendy; Yoshie Harada | Zendy

Open Access

Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA–RuvB protein complex

Author(s) -

Yong-Woon Han,

Tomomi Tani,

Masahito Hayashi,

Takashi Hishida,

Hiroshi Iwasaki,

Hideo Shinagawa,

Yoshie Harada

Publication year - 2006

Publication title -

proceedings of the national academy of sciences

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 5.011

H-Index - 771

eISSN - 1091-6490

pISSN - 0027-8424

DOI - 10.1073/pnas.0600753103

Subject(s) - holliday junction , branch migration , dna , homologous recombination , biophysics , dna repair , rotation (mathematics) , biology , genetics , geometry , mathematics

The Escherichia coli RuvA-RuvB complex promotes branch migration of Holliday junction DNA, which is the central intermediate of homologous recombination. Like many DNA motor proteins, it is suggested that RuvA-RuvB promotes branch migration by driving helical rotation of the DNA. To clarify the RuvA-RuvB-mediated branch migration mechanism in more detail, we observed DNA rotation during Holliday junction branch migration by attaching a bead to one end of cruciform DNA that was fixed to a glass surface at the opposite end. Bead rotation was observed when RuvA, RuvB, and ATP were added to the solution. We measured the rotational rates of the beads caused by RuvA-RuvB-mediated branch migration at various ATP concentrations. The data provided a K(m) value of 65 microM and a V(max) value of 1.6 revolutions per second, which corresponds to 8.3 bp per second. This real-time observation of the DNA rotation not only allows us to measure the kinetics of the RuvA-RuvB-mediated branch migration, but also opens the possibility of elucidating the branch migration mechanism in detail.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research