Amino Acid- and Purine Ribonucleoside-Induced Germination of Bacillus anthracis ΔSterne Endospores: gerS Mediates Responses to Aromatic Ring Structures

Specific combinations of amino acids or purine ribonucleosides and amino acids are required for efficient germination of endospores of Bacillus anthracis ΔSterne, a plasmidless strain, at ligand concentrations in the low-micromolar range. The amino acid l-alanine was the only independent germinant in B. anthracis and then only at concentrations of >10 mM. Inosine and l-alanine both play major roles as cogerminants with several other amino acids acting as efficient cogerminants (His, Pro, Trp, and Tyr combining with l-alanine and Ala, Cys, His, Met, Phe, Pro, Ser, Trp, Tyr, and Val combining with inosine). An ortholog to the B. subtilis tricistronic germination receptor operon gerA was located on the B. anthracis chromosome and named gerS. Disruption of gerS completely eliminated the ability of B. anthracis endospores to respond to amino-acid and inosine-dependent germination responses. The gerS mutation also produced a significant microlag in the aromatic-amino-acid-enhanced-alanine germination pathways. The gerS disruption appeared to specifically affect use of aromatic chemicals as cogerminants with alanine and inosine. We conclude that efficient germination of B. anthracis endospores requires multipartite signals and that gerS-encoded proteins act as an aromatic-responsive germination receptor.