Peptide anchoring spore coat assembly to the outer forespore membrane in Bacillus subtilis.
Spore formation in Bacillus subtilis involves the formation of a thick, proteinaceous shell or coat that is assembled around a specialized membrane known as the outer forespore membrane. Here we present evidence that the assembling coat is tethered to the outer forespore membrane by a 26-amino-acid peptide called SpoVM, which is believed to form an amphipathic helix. We show that proper localization of SpoVM is dependent on SpolVA, a morphogenetic protein that forms the basement layer of the spore coat, and conversely, that proper localization of SpoIVA is dependent on SpoVM. Genetic, biochemical and cytological evidence indicates that this mutual dependence is mediated in part by contact between an amino acid side-chain located near the extreme C-terminus of SpoIVA and an amino acid side-chain on the hydrophilic face of the SpoVM helix. Evidence is also presented that SpoVM adheres to the outer forespore membrane via hydrophobic, amino acid side-chains on the hydrophobic face of the helix. The results suggest that the SpoVM helix is oriented parallel to the membrane with the hydrophobic face buried in the lipid bilayer.
Ramamurthi
Kumaran S
KS
Department of Molecular and Cellular Biology, Harvard University, The Biological Laboratories, 16 Divinity Ave., Cambridge, MA 02138, USA.
Clapham
Katie Rose
KR
Losick
Richard
R
D019943:Amino Acid Substitution; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009154:Mutation; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013171:Spores, Bacterial / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; C084307:spoVM protein, Bacillus subtilis; C030069:spore-specific proteins, Bacillus; D049452:Green Fluorescent Proteins