Functional analysis of the Bacillus subtilis morphogenetic spore coat protein CotE.
Bacterial spores are surrounded by a multilayered proteinaceous shell called the coat. In Bacillus subtilis, a coat protein called CotE guides the assembly of a major subset of coat proteins. To understand how CotE carries out its role in coat morphogenesis, we subjected its gene to mutagenesis and studied the effects of altered versions of CotE on coat formation. We identified regions within the C-terminal 28 amino acids that direct the deposition of the coat proteins CotA, CotB, CotG, CotSA, CotS and 35 kDa and 49 kDa proteins likely to be the spore proteins CotR (formerly known as YvdO) and YaaH respectively. The timing and genetic dependency of CotR accumulation are consistent with control of its gene by sigmaK and GerE. In addition, we identified a 35-amino-acid internal region involved in targeting of CotE to the forespore. Finally, we found that sequences within this 35-amino-acid region as well as within an 18-amino-acid stretch in the N-terminus of CotE direct the formation of CotE multimers, most probably homooligomers. These results suggest that: (i) most interactions between CotE and the coat proteins assembled under CotE control take place at the CotE C-terminus; (ii) an internal region of CotE connects it with the forespore surface; and (iii) interactions between CotE molecules depend on residues within an 18-amino-acid region in the N-terminal half of CotE.
Little
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Department of Microbiology and Immunology, Loyola University Medical Center, 2160 South First Avenue, Maywood, IL 60153, USA.
Driks
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D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D021381:Protein Transport / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012808:Sigma Factor / Q000378:metabolism; D013171:Spores, Bacterial / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; C077494:CotE protein, Bacillus subtilis; D011993:Recombinant Fusion Proteins; D012808:Sigma Factor