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Published online on December 30, 2003, 10.1073/pnas.2533829100
PNAS | January 13, 2004 | vol. 101 | no. 2 | 452-457


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BIOCHEMISTRY
Bacillus subtilis RecU protein cleaves Holliday junctions and anneals single-stranded DNA

Silvia Ayora * {dagger}, Begoña Carrasco *, Ernesto Doncel *, Rudi Lurz {ddagger}, and Juan C. Alonso * §

*Department of Microbial Biotechnology, Centro Nacional de Biotecnología–Consejo Superior de Investigaciones Científicas, Campus Universidad Autónoma de Madrid, 28049 Madrid, Spain; {dagger}Department of Molecular Biology, Universidad Autónoma de Madrid, 28049 Madrid, Spain; and {ddagger}Max-Planck-Institut für molekulare Genetik, Ihnestrasse 73, 14195 Berlin, Germany

Edited by Arthur Landy, Brown University, Providence, RI, and approved October 21, 2003 (received for review June 20, 2003)

Bacillus subtilis RecU protein is involved in homologous recombination, DNA repair, and chromosome segregation. Purified RecU binds preferentially to three- and four-strand junctions when compared to single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) ({approx}10- and {approx}40-fold lower efficiency, respectively). RecU cleaves mobile four-way junctions but fails to cleave a linear dsDNA with a putative cognate site, a finding consistent with a similar genetic defect observed for genes classified within the {epsilon} epistatic group (namely ruvA, recD, and recU). In the presence of Mg2+, RecU also anneals a circular ssDNA and a homologous linear dsDNA with a ssDNA tail and a linear ssDNA and a homologous supercoiled dsDNA substrate. These results suggest that RecU, which cleaves recombination intermediates with high specificity, might also help in their assembly.

DNA repair | RuvC | RuvAB | RusA

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: AGE, agarose gel electrophoresis; dsDNA, double-stranded DNA; EtdBr, ethidium bromide; HJ, Holliday junction; JM, joint molecule; ssDNA, single-stranded DNA.

§ To whom correspondence should be addressed. E-mail: jcalonso{at}cnb.uam.es.


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