ABSTRACT
The activity of the sporulation transcription factor ςE in Bacillus subtilis is governed by an intercellular signal transduction pathway that controls the conversion of the inactive proprotein pro-ςE to the mature and active form of the factor. Here I use immunofluorescence microscopy to show that the activation of the proprotein is associated with its progression through three patterns of subcellular localization. In the predivisional sporangium, pro-ςE was found to be associated with the cytoplasmic membrane. Next, at the stage of asymmetric division, pro-ςE accumulated at the sporulation septum. Finally, after processing, mature ςEwas found to be distributed throughout the mother cell cytoplasm. The results of subcellular fractionation and sedimentation in density gradients of extracts prepared from postdivisional sporangia confirmed that pro-ςE was chiefly present in the membrane fraction and that ςE was predominantly cytoplasmic, findings that suggest that the pro-amino acid sequence is responsible for the sequestration of pro-ςE to the membrane. The results of chemical cross-linking experiments showed that pro-ςE was present in a complex with its putative processing protein, SpoIIGA, or with a protein that depended on SpoIIGA. The membrane association of pro-ςE was, however, independent of SpoIIGA and other proteins specific to B. subtilis. Likewise, accumulation of pro-ςE at the septum did not depend on its interaction with SpoIIGA. Sequestration of pro-ςE to the membrane might serve to facilitate its interaction with SpoIIGA and may be important for preventing its premature association with core RNA polymerase. The implications of these findings for the compartmentalization of ςE are discussed.
Negative regulation of the proteolytic activation of a developmental transcription factor in Bacillus subtilis