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Nucleic Acids Research, 1983, Vol. 11, No. 5 1309-1323
© 1983

MOLECULAR BIOLOGY

Factors involved in the initiation of phage ø29 DNA replication in vitro: requirement of the gene 2 product for the formation of the protein p3-dAMP complex

Luis Blanco, Juan A. Garcia, Miguel A. Peñalva* and Margarita Salas

Centra de Biologia Molecular (CS1C-UAM) Universidad Autónoma Canto Blanco, Madrid 34, Spain

Received January 4, 1983. Accepted January 31, 1983.

To study the requirements for the in vitro formation of the protein p3-dAMP complex, the first step in {varphi}29 DNA replication, extracts from B. subtilis infected with ø29 mutants in genes 2, 3, 5, 6 and 17, involved in DNA synthesis, have been used. The formation of the initiation complex is completely dependent on the presence of a functional gene 2 product, in addition to protein p3 and ø29 DNA-protein p3 as template. ATP is also required, although it can be replaced by other nucleotides. The products of genes 5, 6 and 17 do not seem to be needed in the formation of the initiation complex.

Inhibitors of the host DNA polymerase III, DNA gyrase or RNA polymerase had no effect on the formation of the protein p3-dAMP complex, suggesting that these proteins are not involved in the initiation of ø29 DNA replication. ddATP or aphidicolin, inhibitors of DNA chain elongation, had also no effect on the formation of the initiation complex.

*Present address: Department of Molecular Genetics Antibióticos, S.A., Bravo Murillo 38, Madrid 3, Spain


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L. Blanco and M. Salas
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J. Biol. Chem., April 12, 1996; 271(15): 8509 - 8512.
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