The co-dependence of BxpB/ExsFA and BclA for proper incorporation into the exosporium of Bacillus anthracis.
The outermost layer of the Bacillus anthracis spore consists of an exosporium comprised of two distinct layers, an outer hair-like nap layer and an internal basal layer. The hair-like nap is primarily comprised of the glycosylated collagen-like protein BclA. BclA is found in a trimeric form in close association with many other exosporium proteins in high-molecular weight complexes. We previously had characterized an N-terminal sequence of BclA that is sufficient for incorporation into the exosporium. Here we utilized site-directed mutagenesis to identify BclA residues critical to two steps in this process, positioning of the protein at the site of the developing exosporium basal layer and stable incorporation which includes a proteolytic cleavage of BclA after residue 19. The BxpB (ExsFA) protein is known to be important for proper incorporation of BclA onto the exosporium. BxpB and BclA were found to be expressed at the same time in sporulating cells of B. anthracis and immediately colocalize to high-molecular weight complexes. The BxpB protein was found to be in close proximity to the BclA NTD. BxpB and BclA are co-dependent for exosporium incorporation, with the BclA NTD being sufficient to deliver BxpB to the exosporium.
Department of Veterinary Pathobiology Biochemistry, Bond Life Sciences Center, University of Missouri, Columbia, MO 65211, USA.
Brian M
BM
Thompson
Hsin-Yeh
HY
Hsieh
Krista A
KA
Spreng
George C
GC
Stewart
D001408:Bacillus anthracis / Q000166:cytology / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D015153:Blotting, Western; D005434:Flow Cytometry; D031541:Fluorescence Resonance Energy Transfer; D006023:Glycoproteins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D021381:Protein Transport; D013171:Spores, Bacterial / Q000166:cytology / Q000378:metabolism; D013499:Surface Properties; D013997:Time Factors
D001426:Bacterial Proteins; C470593:BclA protein, Bacillus anthracis; D006023:Glycoproteins; D008562:Membrane Glycoproteins
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