Interactions between partner switcher orthologs BtrW and BtrV regulate type III secretion in Bordetella.
We have recently described a multicomponent cascade that regulates type III secretion in Bordetella. This cascade includes a group of proteins, BtrU, BtrW, and BtrV, that contain an array of domains that define partner-switching complexes previously characterized in gram-positive bacteria. BtrU contains a PP2C-like serine phosphatase domain, BtrW contains a serine kinase/anti-sigma factor motif, and BtrV includes an anti-sigma factor antagonist domain. On the basis of genetic studies and sequence similarity with the RsbU-RsbW-RsbV and SpoIIE-SpoIIAB-SpoIIAA partner switchers of Bacillus subtilis, a series of interactions between Bordetella orthologs have been proposed. Bacterial two-hybrid analysis, tagged protein pull-downs, and in vitro phosphorylation assays were used to characterize interactions between BtrW and BtrV. In addition, BtrV mutants predicted to mimic a constitutively phosphorylated form of BtrV or to be nonphosphorylatable and BtrW mutants defective in serine kinase activity or the ability to bind BtrV were constructed and analyzed. Our results demonstrate that (i) BtrW and BtrV interact with each other, (ii) BtrW phosphorylates BtrV at serine S55, (iii) the conserved serine residue S55 of BtrV plays a key role in BtrV-BtrW interactions, and (iv) the ability of BtrW to phosphorylate BtrV and disrupt BtrV-BtrW binding is essential for the type III secretion process.
Kozak
Natalia A
NA
Department of Microbiology, Immunology and Molecular Genetics, David Geffen School of Medicine at UCLA, 10833 Le Conte Ave., Los Angeles, CA 90095-1747, USA.
Mattoo
Seema
S
Foreman-Wykert
Amy K
AK
Whitelegge
Julian P
JP
Miller
Jeff F
JF
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D016950:Bordetella bronchiseptica / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D012808:Sigma Factor / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; D012808:Sigma Factor; D017346:Protein Serine-Threonine Kinases