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Journal of Bacteriology, August 2004, p. 5450-5459, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5450-5459.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

spoIVH (ykvV), a Requisite Cortex Formation Gene, Is Expressed in Both Sporulating Compartments of Bacillus subtilis

International Environmental and Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu, Tokyo 183-8509,¹ Graduate School of Information Science, Nara Institute of Sciences and Technology, Ikoma, Nara 630-0101,² Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Ueda, Nagano 386-8567, Japan³

Received 27 January 2004/ Accepted 18 May 2004

It is well known that the ykvU-ykvV operon is under the regulation of the ^E-associated RNA polymerase (E^E). In our study, we observed that ykvV is transcribed together with the upstream ykvU gene by E^E in the mother cell and monocistronically under E^G control in the forespore. Interestingly, alternatively expressed ykvV in either the forespore or the mother cell increased the sporulation efficiency in the ykvV background. Studies show that the YkvV protein is a member of the thioredoxin superfamily and also contains a putative Sec-type secretion signal at the N terminus. We observed efficient sporulation in a mutant strain obtained by replacing the putative signal peptide of YkvV with the secretion signal sequence of SleB, indicating that the putative signal sequence is essential for spore formation. These results suggest that YkvV is capable of being transported by the putative Sec-type signal sequence into the space between the double membranes surrounding the forespore. The ability of ykvV expression in either compartment to complement is indeed intriguing and further introduces a new dimension to the genetics of B. subtilis spore formation. Furthermore, electron microscopic observation revealed a defective cortex in the ykvV disruptant. In addition, the expression levels of ^K-directed genes significantly decreased despite normal ^G activity in the ykvV mutant. However, immunoblotting with the anti-^K antibody showed that pro-^K was normally processed in the ykvV mutant, indicating that YkvV plays an important role in cortex formation, consistent with recent reports. We therefore propose that ykvV should be renamed spoIVH.

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