MULTIPLE INTERACTIONS OF A DNA-BINDING PROTEIN IN VIVO. III. PHAGE T4 GENE-32 MUTATIONS DIFFERENTIALLY AFFECT INSERTION-TYPE RECOMBINATION AND MEMBRANE PROPERTIES

Gisela Mosig ¹, William Berquist ¹, and Susan Bock ¹

¹ Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235

We have investigated the in vivo roles of T4 gene- 32 protein in recombination. We have studied the effects of gene- 32 mutations under conditions that allow normal DNA replication and are permissive for progeny production. Under these conditions, certain gene- 32 mutations specifically reduce insertion-type (short-interval) recombination but none affect crossover-type (long-interval) recombination (see Figure 5). Heterozygote frequencies in all gene-32 mutants are similar to or higher than in a gene-32⁺ background and are not correlated with recombination deficiencies. "Recombination-deficient" alleles are dominant or codominant over the "recombination-proficient" gene-32 mutation tsL171. This explains apparent discrepancies between a gene-32 map deduced from two-factor crosses and the map derived from three-factor crosses.

We have also found that the "recombination proficient" mutation tsL171 and its homoalleles suppress the characteristic plaque morphology of rII mutants. Under restrictive conditions, tsL171 is partially suppressed by rII mutations, which allow the use of host ligase in recombination.

Our present and previous results are discussed in terms of current recombination models. We conclude that gene-32 protein functions in recombination by forming a complex with DNA, with recombination enzymes and with membrane components. Since gene-32 protein interacts with many components of this recombination complex, gene-32 mutations may differentially affect various recombination steps.

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