Abstract
Abstract Teichoic acid synthesis was examined in two types of phage-resistant mutants of Staphylococcus aureus H that have altered cell wall composition. In type I mutants (52A2, 52A4, 52B2) which lack N-acetylglucosamine (GlcNAc) on the cell wall ribitol teichoic acid, the CDP-ribitol:acceptor phosphoribitoltransferase was normal but no UDP-GlcNAc:poly(ribitol phosphate) GlcNAc-transferase activity was detected. Revertants of type I mutants recovered the normal levels of GlcNAc in the teichoic acid as well as GlcNAc-transferase activity. Type II mutants (52A5, 52A7, M3, M9) lacked ribitol teichoic acid in the cell wall and no ribitol phosphate polymer was detected in any other cell fraction or in the spent medium. Despite the lack of ribitol teichoic acid, normal activities in vitro of phosphoribitoltransferase and GlcNAc-transferase were found in these mutants. Compared to the parent strain, trichloracetic acid extracts of type II mutants contained high levels of CDP-ribitol. The lack of ribitol teichoic acid in the cell wall is caused presumably by some defect in the membrane or in some unknown factor required in the polymerization or attachment step of the teichoic acid to murein.
Highlights
Type II mutants (5285, 5287, M3, M9) lacked ribitol teichoic acid in the cell wall and no ribitol phosphate polymer was detected in any other cell fraction or in the spent medium
Despite the lack of ribitol teichoic acid, normal activities in vitro of phosphoribitoltransferase and GlcNAc-transferase were found in these mutants
The present paper describes the activities of teichoic acidsynthesizing enzymes in phage-resistant mutants and revertants
Summary
AND MATERIALSGrowth of Bacteria-The isolation and maintenance of the streptomycin-resistant parent strain, X. aureus H(SmR), and of the two types of phage-resistant mutant strains, type I (strains52A2, 52A4, 52B2) and type II (strains 52A5, 52A7, M3, and MS), as well as the isolation of revertants from type II mutants, Ribitol Teichoic Acid Synthesis in Phage-resistant Mutants Vol 245, No 19were described previously [4, 6]. Growth of Bacteria-The isolation and maintenance of the streptomycin-resistant parent strain, X. aureus H(SmR), and of the two types of phage-resistant mutant strains, type I 52A2, 52A4, 52B2) and type II (strains 52A5, 52A7, M3, and MS), as well as the isolation of revertants from type II mutants, Ribitol Teichoic Acid Synthesis in Phage-resistant Mutants Vol 245, No 19. Strains were grown in 0.5% phytone (Baltimore Biological Laboratories), 0.5% yeast extract (Difco), 0.3 % K2HP0,, and 0.2% glucose (pH 7.2) (PYK medium) in a rotary shaker at. In all growth experiments 3 to 5% of a 13- to 15-hour culture was used as an inoculum. Growth was monitored by measuring the optical density of the culture at 585 mp. Isolation of Reuertants-Wild type phage-sensitive revertants from type I phage-resistant mutants were obtained as follows
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