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J Bacteriol, April 1998, p. 2125-2132, Vol. 180, No. 8
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Visualization of Penicillin-Binding Proteins during Sporulation of Streptomyces griseus

Jiang Hao and Kathleen E. Kendrick*

Department of Microbiology, Ohio State University, Columbus, Ohio 43210

Received 10 November 1997/Accepted 2 February 1998

We used fluorescein-tagged beta -lactam antibiotics to visualize penicillin-binding proteins (PBPs) in sporulating cultures of Streptomyces griseus. Six PBPs were identified in membranes prepared from growing and sporulating cultures. The binding activity of an 85-kDa PBP increased fourfold by 10 to 12 h of sporulation, at which time the sporulation septa were formed. Cefoxitin inhibited the interaction of the fluorescein-tagged antibiotics with the 85-kDa PBP and also prevented septum formation during sporulation but not during vegetative growth. The 85-kDa PBP, which was the predominant PBP in membranes of cells that were undergoing septation, preferentially bound fluorescein-6-aminopenicillanic acid (Flu-APA). Fluorescence microscopy showed that the sporulation septa were specifically labeled by Flu-APA; this interaction was blocked by prior exposure of the cells to cefoxitin at a concentration that interfered with septation. We hypothesize that the 85-kDa PBP is involved in septum formation during sporulation of S. griseus.

* Corresponding author. Mailing address: Department of Microbiology, 484 West 12th Ave., Ohio State University, Columbus, OH 43210. Phone: (614) 292-1440. Fax: (614) 292-9195 or (614) 292-8120. E-mail: kendrick.1{at}osu.edu.




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