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Nucleic Acids Research, 1990, Vol. 18, No. 22 6553-6557
© 1990

MOLECULAR BIOLOGY

Characterization of a DNA binding protein of bacteriophage PRD1 involved in DNA replication

Tiina M. Pakula, Javier Caldentey, Manuel Serrano1, Crisanto Gutierrez1, Jose M. Hermoso1, Margarita Salas1 and Dennis H. Bamford*,

Department of Genetics, University of Helsinki Arkadiankatu 7, SF-00100 Helsinki, Finland 1Centra de Biologia Molecular (CSIC-UAM) Canto Blanco, 28049 Madrid, Spain

*To whom correspondence should be addressed

Received August 8, 1990. Revised October 15, 1990. Accepted October 15, 1990.

Eschehchia coli phage PRD1 protein P12, involved In PRD1 DNA replication in vivo, has been highly purified from E. coli cells harboring a gene Xll-containing plasmid. Protein P12 binds to single-stranded DNA as shown by gel retardation assays and nuclease protection experiments. Binding of protein P12 to single-stranded DNA increases about 14% the contour length of the DNA as revealed by electron microscopy. Binding to single-stranded DNA seems to be cooperative, and it is not sequence specific. Protein P12 also binds to double-stranded DNA although with an affinity 10 times lower than to single-stranded DNA. Using the in vitro phage {Phi}29 DNA replication system, it is shown that protein P12 stimulates the overall {Phi}29 DNA replication.


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