Interactions of Bacillus subtilis Basement Spore Coat Layer Proteins

Abstract

Bacillus subtilis endospores are exceptionally resistant cells encircled by two protective layers: a petidoglycan layer, termed the cortex, and the spore coat, a proteinaceous layer. The formation of both structures depends upon the proper assembly of a basement coat layer, which is composed of two proteins, SpoIVA and SpoVM. The present work examines the interactions of SpoIVA and SpoVM with coat proteins recruited to the spore surface during the early stages of coat assembly. We showed that the alanine racemase YncD associates with two morphogenetic proteins, SpoIVA and CotE. Mutant spores lacking the yncD gene were less resistant against wet heat and germinated to a greater extent than wild-type spores in the presence of micromolar concentrations of l-alanine. In seeking a link between the coat and cortex formation, we investigated the interactions between SpoVM and SpoIVA and the proteins essential for cortex synthesis and found that SpoVM interacts with a penicillin-binding protein, SpoVD, and we also demonstrated that SpoVM is crucial for the proper localization of SpoVD. This study shows that direct contacts between coat morphogenetic proteins with a complex of cortex-synthesizing proteins could be one of the tools by which bacteria couple cortex and coat formation.

Oral vaccination of fish against vibriosis using spore-display technology

“…GFP display at the spores’ surface was further confirmed by fluorescence microscopy in both derivatives, and the C-terminal version CotY-H6-GFP exhibited a higher fluorescence signal than the N-terminal H6-GFP-CotY. A possible explanation for the inability to detect the N-terminal fusions by western blotting is the strong and tight interaction network formed by cross-linked protein complexes that are present at the spore outermost layers structure ( 43 , 90 – 92 ). CotY appears to interact with most of the spore crust proteins, including itself.…”

“…CotY appears to interact with most of the spore crust proteins, including itself. Indeed, the strongest functional complexes involving this protein have arisen from self-interactions (82,(90)(91)(92)(93)(94). Moreover, considerably strong bonds between CotY-CotZ and CotY-CotV have been frequently observed (90)(91)(92).…”

“…Indeed, the strongest functional complexes involving this protein have arisen from self-interactions (82,(90)(91)(92)(93)(94). Moreover, considerably strong bonds between CotY-CotZ and CotY-CotV have been frequently observed (90)(91)(92). These protein complexes may be resistant to the degree of denaturation performed, and thus unlikely to run in the polyacrylamide gel.…”

Assessment of temperature and acid tolerance of Bacillus subtilis isolated from a Brazilian fruit juice-added soy beverage

“…subtillis strain PY79 submitted to 100°C and 110°C exhibited D-values of 8 min and approximately 0.7 min, respectively (19) . Bacillus atrophaeus spores were submitted to heat treatment at 105°C and exhibited a D-value of 25.6 min (±3.2min) (20) .…”

Special Issue “Bacillus subtilis as a Model Organism to Study Basic Cell Processes”

“…More than 80 coat proteins are localized into four distinct morphological layers. Krajčíková et al [8] investigated the interactions between SpoVM and SpoIVA and the proteins essential for cortex synthesis. They found and characterized their protein partners localized in the basement spore coat layer.…”