A Polypeptide Bacteriophage Receptor: Modified Cell Wall Protein Subunits in Bacteriophage-Resistant Mutants of Bacillus sphaericus Strain P-1

Abstract

Bacillus sphaericus strain P-1 has previously been shown to have a tetragonally arrayed (T layer) protein which forms the outer layer of the cell wall. The T layer was quantitatively extracted from whole cells by 6 M urea, and the T layer subunits were purified by electrophoresis of the extracts on acrylamide gels containing 0.1% sodium dodecyl sulfate or 6 M urea. Using ethylene diacrylate cross-linked gels, the T layer was found to make up 16% of the total cellular protein. A virulent bacteriophage which is inactivated by purified T layer was isolated from soil. Twenty-four phage-resistant mutants were isolated, of which 17 had T layer subunits of increased mobility on sodium dodecyl sulfate acrylamide gels. No mutants devoid of T layer were found. Mutants were grouped into six classes according to the molecular weight of their T layer subunits. These ranged from that of the wild type, 150,000 down to 86,000. Two mutants from different classes were examined in detail. Cells of the mutant strains did not adsorb phage nor did cell walls isolated from these mutants inactivate phage. The amino acid composition of the T layers from mutants differed little from that of the wild-type T layer.