Abstract
The Bacillus subtilis Rap family of proteins are characterized by protein-protein interaction modules containing the so-called tetratricopeptide repeats (TPRs). The six TPR motifs of RapC mediate its interaction with the pentapeptide inhibitor PhrC (ERGMT) or with its target protein ComA, a phosphorylation-dependent response regulator transcription factor for genetic competence. Our results show that RapC interaction with ComA inhibits the response regulator's ability to bind its target DNA promoter but does not affect its phosphorylation state. RapC binds equally well to ComA or to ComA approximately P. The PhrC pentapeptide binds to RapC and inhibits its interaction with ComA. The D195 residue in TPR3 and the P263 residue in TPR5 of RapC are critical for the interaction with PhrC as their mutation to asparagine or leucine, respectively, prevents peptide inhibitory activity. The RapC mechanism of regulating ComA activity is a new example of how TPR motifs and their structural organization have been adapted for different specific functions within the B. subtilis Rap family.
Publication types
-
Research Support, Non-U.S. Gov't
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Amino Acid Motifs / physiology
-
Amino Acid Sequence
-
Autoradiography
-
Bacillus subtilis / genetics
-
Bacillus subtilis / metabolism*
-
Bacterial Proteins / chemistry
-
Bacterial Proteins / genetics
-
Bacterial Proteins / isolation & purification
-
Bacterial Proteins / metabolism*
-
DNA, Bacterial / metabolism*
-
DNA-Binding Proteins / chemistry
-
DNA-Binding Proteins / genetics
-
DNA-Binding Proteins / isolation & purification
-
DNA-Binding Proteins / metabolism*
-
Electrophoresis, Polyacrylamide Gel
-
Electrophoretic Mobility Shift Assay
-
Esterases / chemistry
-
Esterases / genetics
-
Esterases / isolation & purification
-
Esterases / metabolism*
-
Gene Expression Regulation, Bacterial
-
Genes, Reporter
-
Molecular Sequence Data
-
Mutagenesis, Site-Directed
-
Operon
-
Phosphoprotein Phosphatases / metabolism
-
Promoter Regions, Genetic
-
Protein Binding
-
Protein Structure, Tertiary
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / metabolism
-
Repressor Proteins / chemistry
-
Repressor Proteins / genetics
-
Repressor Proteins / metabolism
-
Signal Transduction*
-
beta-Galactosidase / genetics
-
beta-Galactosidase / metabolism
Substances
-
Bacterial Proteins
-
ComA protein, Bacteria
-
DNA, Bacterial
-
DNA-Binding Proteins
-
PhrC protein, Bacillus subtilis
-
Recombinant Fusion Proteins
-
Repressor Proteins
-
Esterases
-
RapC protein, Bacillus subtilis
-
Phosphoprotein Phosphatases
-
beta-Galactosidase