Characterization of the Bacillus subtilis spore morphogenetic coat protein CotO.
Bacillus spores are protected by a structurally and biochemically complex protein shell composed of over 50 polypeptide species, called the coat. Coat assembly in Bacillus subtilis serves as a relatively tractable model for the study of the formation of more complex macromolecular structures and organelles. It is also a critical model for the discovery of strategies to decontaminate B. anthracis spores. In B. subtilis, a subset of coat proteins is known to have important roles in assembly. Here we show that the recently identified B. subtilis coat protein CotO (YjbX) has an especially important morphogenetic role. We used electron and atomic force microscopy to show that CotO controls assembly of the coat layers and coat surface topography as well as biochemical and cell-biological analyses to identify coat proteins whose assembly is CotO dependent. cotO spores are defective in germination and partially sensitive to lysozyme. As a whole, these phenotypes resemble those resulting from a mutation in the coat protein gene cotH. Nonetheless, the roles of CotH and CotO and the proteins whose assembly they direct are not identical. Based on fluorescence and electron microscopy, we suggest that CotO resides in the outer coat (although not on the coat surface). We propose that CotO and CotH participate in a late phase of coat assembly. We further speculate that an important role of these proteins is ensuring that polymerization of the outer coat layers occurs in such a manner that contiguous shells, and not unproductive aggregates, are formed.
McPherson
D C
DC
Department of Microbiology and Immunology, Loyola University Medical Center, 2160 S. 1st Ave., Maywood, IL 60153, USA.
Kim
H
H
Hahn
M
M
Wang
R
R
Grabowski
P
P
Eichenberger
P
P
Driks
A
A
D001412:Bacillus subtilis / Q000502:physiology*; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D049452:Green Fluorescent Proteins / Q000032:analysis; D046911:Macromolecular Substances / Q000378:metabolism; D018625:Microscopy, Atomic Force; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009024:Morphogenesis; D016254:Mutagenesis, Insertional; D013171:Spores, Bacterial / Q000737:chemistry / Q000648:ultrastructure*
D001426:Bacterial Proteins; C101016:CotH protein, Bacillus subtilis; D046911:Macromolecular Substances; D049452:Green Fluorescent Proteins