Multiple interactions of a DNA-binding protein in vivo. II. Effects of host mutations on DNA replication of phage T4 gene 32 mutants

Breschkin, A.M.; Mosig, G.

Journal of Molecular Biology 112(2): 295-308


ISSN/ISBN: 0022-2836
PMID: 327075
DOI: 10.1016/s0022-2836(77)80145-9
Accession: 068526331

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T4 gene 32 protein specifically interacts with T4 DNA polymerase and stimulates its polymerizing activity in vitro. To determine which DNA metabolism steps require this interaction in vivo studies were done to find out in which steps host functions might substitute for a mutationally altered gene 32 protein and whether T4 or Escherichia coli DNA polymerases are required under these conditions in initiation of DNA replication, in recombination and/or in DNA elongation. Several host functions (e.g., dnaC, dnaG) probably acting in a complex, can substitute for tene 32 protein in the 1st round of T4 DNA replication (primary replication). This replication requires functional T4 DNA polymerase and dose not depend on E. coli polymerase III. Specific interaction of 32 protein and T4 DNA polymerase is important in initiation of DNA replication and in recombination. E. coli DNA polymerase I can partially substitute for the latter function of the gene 32 T4 DNA polymerase complex. Gene 32 protein, besides binding to DNA, apparently interacts with T4 DNA polymerase, ligase and recombination nucleases and facilitates the coordinated action of these enzymes during recombination. Interactions of gene 32 protein with other proteins during DNA elongation must be different from interactions seen in recombination and initiation of DNA replication.