We have studied the role of the red and gam genes in lambda replication, after infection of wild type and two recombination deficient hosts. Our results show that the rate of phage DNA replication is abnormally low in the absence of red function, in rec+ as well as rec- (A- and A-B-) bacteria. It appears that the virus general recombination proteins play some role in lambda replication that cannot be assumed by the general recombination proteins of its bacterial host. The red- defect in replication results in a decrease in the total amount of intracellular phage DNA. This DNA, nevertheless, seems normal in structure and is matured and packaged with good efficiency. In rec+ and recA- hosts infected with gam- mutants, the rate of lambda replication is also low, but in this case, abnormal DNA structures are produced at late times. The gam mutation seems to alter the program of replication such that circular molecules are produced not only at early times, but continuously, throughout the lytic cycle. This, and other facts, suggest that the gam protein is required for the transition from "early" to "late" replication. This requirement for gam function is not observed in recA-B- hosts, in which gam mutants replicate at a normal rate and produce DNA indistinguishable from that made by wild type phage. Thus, the gam requirement seems to involve an interaction of this phage protein with the product of the host's recB gene. Other evidence for such interaction comes from our finding that, in vivo, the gam protein does inhibit presumed action of the host's BC nuclease. In the gam- mutant infections, which are blocked in late replication, absence of a general recombination system seems to create a severe defect in maturation of intracellular phage DNA. This defect, unlike the one affecting λ replication rate, can be alleviated by either the red or rec functions and is correlated with the inability of the mutant phages to make DNA concatemers. Since other late functions (i.e. late messenger RNA production) appear to be normal, we conclude that concatemer formation, via replication or recombination, is an essential step in phage development.
All Science Journal Classification (ASJC) codes
- Structural Biology
- Molecular Biology