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Spore Coat Architecture of Clostridium novyi NT Spores

Department of Chemistry, Materials and Life Sciences, Lawrence Livermore National Laboratory, L-234, Livermore, California 94551,¹ Department of Biology, Johns Hopkins University, 3400 N. Charles St., Baltimore, Maryland 21218,² The Howard Hughes Medical Institute and the Ludwig Center for Cancer Genetics and Therapeutics at The Johns Hopkins Sidney Kimmel Comprehensive Cancer Center, 1650 Orleans Street, CRB1, Baltimore, Maryland 21231³

Received 14 May 2007/ Accepted 14 June 2007

Spores of the anaerobic bacterium Clostridium novyi NT are able to germinate in and destroy hypoxic regions of tumors in experimental animals. Future progress in this area will benefit from a better understanding of the germination and outgrowth processes that are essential for the tumorilytic properties of these spores. Toward this end, we have used both transmission electron microscopy and atomic force microscopy to determine the structure of both dormant and germinating spores. We found that the spores are surrounded by an amorphous layer intertwined with honeycomb parasporal layers. Moreover, the spore coat layers had apparently self-assembled, and this assembly was likely to be governed by crystal growth principles. During germination and outgrowth, the honeycomb layers, as well as the underlying spore coat and undercoat layers, sequentially dissolved until the vegetative cell was released. In addition to their implications for understanding the biology of C. novyi NT, these studies document the presence of proteinaceous growth spirals in a biological organism.

Published ahead of print on 22 June 2007.