Nucleotide Cofactor-Dependent Structural Change of Xenopus laevis Rad51 Protein Filament Detected by Small-Angle Neutron Scattering Measurements in Solution

Rad51 protein, a eukaryotic homologue of RecA protein, forms a filamentous complex with DNA and catalyzes homologous recombination. We have analyzed the structure of Xenopus Rad51 protein (XRad51.1) in solution by small-angle neutron scattering (SANS). The measurements showed that XRad51.1 forms a helical filament independently of DNA. The sizes of the cross-sectional and helical pitch of the filament could be determined, respectively, from a Guinier plot and the position of the subsidiary maximum of SANS data. We observed that the helical structure is modified by nucleotide binding as in the case of RecA. Upon ATP binding under high-salt conditions (600 mM NaCl), the helical pitch of XRad51.1 filament was increased from 8 to 10 nm and the cross-sectional diameter decreased from 7 to 6 nm. The pitch sizes of XRad51.1 are similar to, though slightly larger than, those of RecA filament under corresponding conditions. A similar helical pitch size was observed by electron microscopy for budding yeast Rad51 [Ogawa, T., et al. (1993) Science 259, 1896-1899]. In contrast to the RecA filament, the structure of XRad51.1 filament with ADP is not significantly different from that with ATP. Thus, the hydrolysis of ATP to ADP does not modify the helical filament of XRad51.1. Together with our recent observation that ADP does not weaken the XRad51.1/DNA interaction, the effect of ATP hydrolysis on XRad51.1 nucleofilament should be very different from that on RecA.

Kim, Hye-Kyung,1970Chalmers tekniska högskola,Chalmers University of Technology (author)
Maeshima, K. (author)
Tuite, Eimer,1966Chalmers tekniska högskola,Chalmers University of Technology (author)
Morimatsu, K. (author)
Horii, T. (author)
Mortensen, K. (author)
Nordén, Bengt,1945Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)norden (author)
Takahashi, M. (author)
Chalmers tekniska högskola (creator_code:org_t)

In:Biochemistry: American Chemical Society (ACS)36:44, s. 13524-135291520-49950006-2960

By the author/editor: Ellouze, C.; Kim, Hye-Kyung, ...; Maeshima, K.; Tuite, Eimer, 19 ...; Morimatsu, K.; Horii, T.; show more...; Mortensen, K.; Nordén, Bengt, 1 ...; Takahashi, M.; show less...

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NATURAL SCIENCES: NATURAL SCIENCES; and Chemical Science ...; and Physical Chemist ...

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