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Journal of Bacteriology, September 2006, p. 6652-6660, Vol. 188, No. 18
0021-9193/06/$08.00+0     doi:10.1128/JB.00391-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Granular Layer in the Periplasmic Space of Gram-Positive Bacteria and Fine Structures of Enterococcus gallinarum and Streptococcus gordonii Septa Revealed by Cryo-Electron Microscopy of Vitreous Sections

Benoît Zuber,^1* Marisa Haenni,² Tânia Ribeiro,³ Kathrin Minnig,^2, Fátima Lopes,³ Philippe Moreillon,² and Jacques Dubochet¹

Laboratory of Ultrastructural Analysis, Biophore Building, University of Lausanne, CH-1015 Lausanne, Switzerland,¹ Department of Fundamental Microbiology, Biophore Building, University of Lausanne, CH-1015 Lausanne, Switzerland,² IBET/ITQB, Laboratory of Stress by Antibiotics and Virulence of Enterococci, Apartado 12, 2781-901 Oeiras, Portugal³

Received 20 March 2006/ Accepted 4 July 2006

High-resolution structural information on optimally preserved bacterial cells can be obtained with cryo-electron microscopy of vitreous sections. With the help of this technique, the existence of a periplasmic space between the plasma membrane and the thick peptidoglycan layer of the gram-positive bacteria Bacillus subtilis and Staphylococcus aureus was recently shown. This raises questions about the mode of polymerization of peptidoglycan. In the present study, we report the structure of the cell envelope of three gram-positive bacteria (B. subtilis, Streptococcus gordonii, and Enterococcus gallinarum). In the three cases, a previously undescribed granular layer adjacent to the plasma membrane is found in the periplasmic space. In order to better understand how nascent peptidoglycan is incorporated into the mature peptidoglycan, we investigated cellular regions known to represent the sites of cell wall production. Each of these sites possesses a specific structure. We propose a hypothetic model of peptidoglycan polymerization that accommodates these differences: peptidoglycan precursors could be exported from the cytoplasm to the periplasmic space, where they could diffuse until they would interact with the interface between the granular layer and the thick peptidoglycan layer. They could then polymerize with mature peptidoglycan. We report cytoplasmic structures at the E. gallinarum septum that could be interpreted as cytoskeletal elements driving cell division (FtsZ ring). Although immunoelectron microscopy and fluorescence microscopy studies have demonstrated the septal and cytoplasmic localization of FtsZ, direct visualization of in situ FtsZ filaments has not been obtained in any electron microscopy study of fixed and dehydrated bacteria.

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* Corresponding author. Mailing address: Laboratory of Ultrastructural Analysis, Biophore Building, University of Lausanne, CH-1015 Lausanne, Switzerland. Phone: 41 21 6924289. Fax: 41 21 6924285. E-mail: Benoit.Zuber{at}unil.ch.

Present address: ZLB Behring AG, Wankdorfstrasse 10, CH-3000 Bern, Switzerland.

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Journal of Bacteriology, September 2006, p. 6652-6660, Vol. 188, No. 18
0021-9193/06/$08.00+0     doi:10.1128/JB.00391-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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