Localization of the transglutaminase cross-linking sites in the Bacillus subtilis spore coat protein GerQ.
The Bacillus subtilis spore coat protein GerQ is necessary for the proper localization of CwlJ, an enzyme important in the hydrolysis of the peptidoglycan cortex during spore germination. GerQ is cross-linked into high-molecular-mass complexes in the spore coat late in sporulation, and this cross-linking is largely due to a transglutaminase. This enzyme forms an epsilon-(gamma-glutamyl) lysine isopeptide bond between a lysine donor from one protein and a glutamine acceptor from another protein. In the current work, we have identified the residues in GerQ that are essential for transglutaminase-mediated cross-linking. We show that GerQ is a lysine donor and that any one of three lysine residues near the amino terminus of the protein (K2, K4, or K5) is necessary to form cross-links with binding partners in the spore coat. This leads to the conclusion that all Tgl-dependent GerQ cross-linking takes place via these three lysine residues. However, while the presence of any of these three lysine residues is essential for GerQ cross-linking, they are not essential for the function of GerQ in CwlJ localization.
Monroe
Alicia
A
Department of Molecular, Microbial, and Structural Biology, University of Connecticut Health Center, Farmington, 06030, USA.
Setlow
Peter
P
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001704:Biopolymers / Q000378:metabolism; D015153:Blotting, Western; D005973:Glutamine / Q000378:metabolism; D006867:Hydrolases / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016415:Sequence Alignment; D013171:Spores, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism
D001426:Bacterial Proteins; D001704:Biopolymers; C030069:spore-specific proteins, Bacillus; D005973:Glutamine; D011503:Transglutaminases; C433796:CwlJ protein, Bacillus subtilis; D006867:Hydrolases; D008239:Lysine