Coupling of σG Activation to Completion of Engulfment During Sporulation of Bacillus Subtilis Survives Large Perturbations to DNA Translocation and Replication

Spore formation in Bacillus subtilis is characterized by activation of RNA polymerase sigma factors, including the late-expressed σ(G). During spore formation an asymmetric division occurs, yielding the smaller prespore and the larger mother cell. At division, only 30% of the chromosome is in the prespore, and the rest is then translocated into the prespore. Following completion of engulfment of the prespore by the mother cell, σ(G) is activated in the prespore. Here we tested the link between engulfment and σ(G) activation by perturbing DNA translocation and replication, which are completed before engulfment. One approach was to have large DNA insertions in the chromosome; the second was to have an impaired DNA translocase; the third was to use a strain in which the site of termination of chromosome replication was relocated. Insertion of 2.3 Mb of Synechocystis DNA into the B. subtilis genome had the largest effect, delaying engulfment by at least 90 min. Chromosome translocation was also delayed and was completed shortly before the completion of engulfment. Despite the delay, σ(G) became active only after the completion of engulfment. All results are consistent with a strong link between completion of engulfment and σ(G) activation. They support a link between completion of chromosome translocation and completion of engulfment.