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Journal of Bacteriology, December 1998, p. 6493-6502, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Analysis of Outgrowth of Bacillus subtilis Spores Lacking Penicillin-Binding Protein 2a

Thomas Murray,1 David L. Popham,2 Christine B. Pearson,3 Arthur R. Hand,3,4 and Peter Setlow1,*

Department of Biochemistry,1 Central Electron Microscope Facility,3 and Department of Pediatric Dentistry,4 University of Connecticut Health Center, Farmington, Connecticut 06032, and Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 240612

Received 27 July 1998/Accepted 15 October 1998

The loss of Bacillus subtilis penicillin-binding protein (PBP) 2a, encoded by pbpA, was previously shown to slow spore outgrowth and result in an increased diameter of the outgrowing spore. Further analyses to define the defect in pbpA spore outgrowth have shown that (i) outgrowing pbpA spores exhibited only a slight defect in the rate of peptidoglycan (PG) synthesis compared to wild-type spores, but PG turnover was significantly slowed during outgrowth of pbpA spores; (ii) there was no difference in the location of PG synthesis in outgrowing wild-type and pbpA spores once cell elongation had been initiated; (iii) outgrowth and elongation of pbpA spores were dramatically affected by the levels of monovalent or divalent cations in the medium; (iv) there was a partial redundancy of function between PBP2a and PBP1 or -4 during spore outgrowth; and (v) there was no difference in the structure of PG from outgrowing wild-type spores or spores lacking PBP2a or PBP2a and -4; but also (vi) PG from outgrowing spores lacking PBP1 and -2a had transiently decreased cross-linking compared to PG from outgrowing wild-type spores, possibly due to the loss of transpeptidase activity.

* Corresponding author. Mailing address: Department of Biochemistry, University of Connecticut Health Center, Farmington, CT 06032. Phone: (203) 679-2607. Fax: (203) 679-3408. E-mail: setlow{at}sun.uchc.edu.

Journal of Bacteriology, December 1998, p. 6493-6502, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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