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Bacteriophage PMB12 conversion of the sporulation defect in RNA polymerase mutants of Bacillus subtilis
Abstract:
The pseudotemperate phage PMB12 was isolated from soil on the basis of its ability to enhance the rate of sporulation of Bacillus subtilis 168. PMB12 was subsequently shown to convert the sporulation defect in two genetically distinct classes of sporulation mutants. One class includes those rifampin-resistant mutants that are also spore-negative (mutated at the rif locus). The other class includes a strain carrying the sporulation mutation spoCM-1. The spoCM-1 mutation is linked to cysA15 by PBS1 transduction but is distinct from the rif locus. Several other sporulation mutants were not converted by PMB12. PMB12 is related to phage PBS1. However, PBS1 did not convert the above sporulation mutants. The replication of PBS2, a clear-plaquing derivative of PBS1, is rifampin insensitive, apparently due to a phage-induced rifampin-insensitive RNA polymerase. PMB12 replication is also rifampin insensitive.
Insights
The pseudotemperate phage PMB12 enhances Bacillus subtilis sporulation by correcting specific genetic defects. This phage, related to PBS1, shows unique conversion capabilities in certain sporulation mutants.
Area of Science:
- Microbiology
- Bacteriology
- Molecular Biology
Background:
- Bacillus subtilis sporulation is a complex developmental process.
- Specific mutations can lead to sporulation defects.
- Bacteriophages can interact with bacterial genetic processes.
Purpose of the Study:
- To investigate the ability of pseudotemperate phage PMB12 to enhance Bacillus subtilis sporulation.
- To characterize the genetic basis of PMB12's effect on sporulation mutants.
- To compare PMB12 with related phages like PBS1.
Main Methods:
- Isolation of phage PMB12 from soil.
- Testing PMB12's effect on various Bacillus subtilis sporulation mutants.
- Genetic mapping of spoCM-1 mutation using PBS1 transduction.
- Assessing rifampin resistance of phage replication.
Main Results:
- Phage PMB12 enhanced sporulation in Bacillus subtilis 168.
- PMB12 corrected sporulation defects in rifampin-resistant mutants (rif locus) and spoCM-1 mutants.
- The spoCM-1 mutation is distinct from the rif locus.
- Phage PMB12 and its relative PBS1 exhibit different conversion capabilities.
- PMB12 replication is rifampin-insensitive.
Conclusions:
- Pseudotemperate phage PMB12 possesses unique properties for correcting specific Bacillus subtilis sporulation mutations.
- PMB12's ability to convert certain mutants highlights its potential as a tool in genetic studies.
- The rifampin-insensitive replication of PMB12 suggests a novel mechanism, possibly involving phage-induced RNA polymerase.