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J. Biol. Chem., Vol. 277, Issue 39, 35969-35979, September 27, 2002
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From the Genetic evidence suggests that the SPP1-encoded
gene 35 product (G35P) is essential for phage
DNA replication. Purified G35P binds single-strand DNA
(ssDNA) and double-strand (dsDNA) and specifically interacts with
SPP1-encoded replicative DNA helicase G40P and SSB protein
G36P. G35P promotes joint molecule formation between a circular ssDNA and a homologous linear dsDNA with an ssDNA
tail. Joint molecule formation requires a metal ion but is independent
of a nucleotide cofactor. Joint molecules formed during these reactions
contain a displaced linear ssDNA strand. Electron microscopic analysis
shows that G35P forms a multimeric ring structure in ssDNA
tails of dsDNA molecules and left-handed filaments on ssDNA.
G35P promotes strand annealing at the AT-rich region of
SPP1 oriL on a supercoiled template. These results
altogether are consistent with the hypothesis that the homologous
pairing catalyzed by G35P is an integral part of SPP1 DNA
replication. The loading of G40P at a D-loop
(ori DNA or at any stalled replication fork) by
G35P could lead to replication fork reactivation.
Homologous-pairing Activity of the Bacillus subtilis
Bacteriophage SPP1 Replication Protein G35P*
§,,,
,, and**
Departmento de Biotecnología
Microbiana, Centro Nacional de Biotecnología, Consejo Superior
de Investigaciones Científicas, Campus Universidad
Autónoma de Madrid, Madrid 28049, Spain, the
§ Departamento de Biología Molecular, Universidad
Autónoma de Madrid, 28049 Madrid, Spain, the
¶ Max-Planck-Institut für molekulare Genetik, Ihnestrasse
73, D-14195 Berlin, Germany, and the Department of
Biochemistry and Molecular Genetics, University of Virginia,
Charlottesville, Virginia 22908
*
This work was supported by Grants BMC2000-0548 and
BIO2001-4342-E from Dirección General de
Investigación-Ministerio de Ciencia y Tecnología
(DGI-MCYT) and QLRT-2000-00365 from the European Union (to
J. C. A.).The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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