The coat morphogenetic protein SpoVID is necessary for spore encasement in Bacillus subtilis.
Endospores formed by Bacillus subtilis are encased in a tough protein shell known as the coat, which consists of at least 70 different proteins. We investigated the process of spore coat morphogenesis using a library of 40 coat proteins fused to green fluorescent protein and demonstrate that two successive steps can be distinguished in coat assembly. The first step, initial localization of proteins to the spore surface, is dependent on the coat morphogenetic proteins SpoIVA and SpoVM. The second step, spore encasement, requires a third protein, SpoVID. We show that in spoVID mutant cells, most coat proteins assembled into a cap at one side of the developing spore but failed to migrate around and encase it. We also found that SpoIVA directly interacts with SpoVID. A domain analysis revealed that the N-terminus of SpoVID is required for encasement and is a structural homologue of a virion protein, whereas the C-terminus is necessary for the interaction with SpoIVA. Thus, SpoVM, SpoIVA and SpoVID are recruited to the spore surface in a concerted manner and form a tripartite machine that drives coat formation and spore encasement.
Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY 10003, USA.
Katherine H
KH
Wang
Anabela L
AL
Isidro
Lia
L
Domingues
Haig A
HA
Eskandarian
Peter T
PT
McKenney
Kevin
K
Drew
Paul
P
Grabowski
Ming-Hsiu
MH
Chua
Samantha N
SN
Barry
Michelle
M
Guan
Richard
R
Bonneau
Adriano O
AO
Henriques
Patrick
P
Eichenberger
D019943:Amino Acid Substitution; D001412:Bacillus subtilis / Q000166:cytology / Q000235:genetics / Q000502:physiology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D017124:Conserved Sequence / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008856:Microscopy, Fluorescence; D009024:Morphogenesis / Q000235:genetics; D009154:Mutation; D019151:Peptide Library; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013171:Spores, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; C080140:spoVID protein, Bacillus subtilis; D049452:Green Fluorescent Proteins