FIG 4.

Bacillus anthracis LCP enzymes attach teichoic acid to Staphylococcus aureus peptidoglycan. (A) Purified murein sacculi from S. aureus MSSA1112 (WT with empty vector) grown in the absence (lane 1) or presence (lane 2) of tunicamycin (tun) were treated with dilute sodium hydroxide and the released wall teichoic acids (WTA) were separated by PAGE and stained with alcian blue and silver. Lanes 3 to 10, WTA extracts of murein sacculi from the S. aureus Δlcp mutant carrying either empty plasmid (vector), control plasmid with S. aureus lcpA (plcpA_Sa), or plasmids with any one of the six lcp genes of B. anthracis: plcpB1, plcpB2, plcpB3, plcpB4, plcpC, and plcpD. The bottom portion shows an immunoblot for staphylococcal sortase (SrtA) using bacterial lysate samples for the corresponding WTA extracts. (B) Growth of the S. aureus Δlcp mutant carrying vector, plcpA_Sa, or plasmids with the six lcp genes of B. anthracis was monitored by first normalizing stationary-phase cultures to an A₆₀₀ of 5, next diluting these cultures (1:100) into fresh brain heart infusion broth, and then recording the A₆₀₀ during the 12-h incubation at 37°C with rotation.