Subcellular localization of proteins involved in the assembly of the spore coat of Bacillus subtilis

Abstract

Spores of the bacterium Bacillus subtilis are encased in a two-layered protein shell, which consists of an electron-translucent, lamellar inner coat, and an electron-dense outer coat. The coat protein CotE is both a structural component of the coat and a morphogenetic protein that is required for the assembly of the outer coat. We now show that CotE is located in the outer coat of the mature spore and that at an intermediate stage of sporulation, when the developing spore (the forespore) is present as a free protoplast within the sporangium, CotE is localized in a ring that surrounds the forespore but is separated from it by a small gap. We propose that the ring is the site of assembly of the outer coat and that the gap is the site of formation of the inner coat. Assembly of the ring depends on the sporulation protein SpoIVA, which sits close to or on the surface of the outer membrane that encircles the forespore. We propose that SpoIVA creates a basement layer around the forespore on which coat assembly takes place. The subcellular localization and assembly of CotE and other coat proteins are therefore determined by the capacity of SpoIVA to recognize and adhere to a specific surface within the sporangium, the outer membrane of the forespore.