A tyrosine phosphorylation switch controls the interaction between the transmembrane modulator protein Wzd and the tyrosine kinase Wze of Lactobacillus rhamnosus

• Published in: BMC microbiology 2015-02, Vol.15 (1), p.40-40, Article 40
• Main Authors: Kang, Hye-Ji, Gilbert, Christophe, Badeaux, Frédérique, Atlan, Danièle, LaPointe, Gisèle
• Format: Article
• Language: English
• Subjects: Analysis; Antibodies; Bacterial Proteins - metabolism; Bacteriology; Gene Expression Regulation, Bacterial; Immunology; Lactobacillus rhamnosus - genetics; Lactobacillus rhamnosus - metabolism; Lactobacillus rhamnosus - physiology; Life Sciences; Microbiology and Parasitology; Phosphorylation; Physiological aspects; Protein Binding; Protein Interaction Mapping; Protein kinases; Protein Processing, Post-Translational; Protein-Tyrosine Kinases - metabolism; Viral antibodies; Virology
• ISSN: 1471-2180; 1471-2180
• DOI: 10.1186/s12866-015-0371-2

One proposed mechanism for assembly of secreted heteropolysaccharides by many Gram positive bacteria relies on the coordinated action of a polymerization complex through reversible phosphorylation events. The role of the tyrosine protein kinase transmembrane modulator is, however, not well understood. The protein sequences deduced from the wzb, wzd and wze genes from Lactobacillus rhamnosus ATCC 9595 and RW-9595 M contain motifs also found in corresponding proteins CpsB, CpsC and CpsD from Streptococcus pneumoniae D39 (serotype 2). Use of an anti-phosphotyrosine antibody demonstrated that both Wzd and Wze can be found in tyrosine phosphorylated form. When tyrosine 266 was mutated to phenylalanine, WzdY266F showed slightly less phosphorylated protein than those produced by using eight other tyrosine mutated Wzd genes, when expressed along with Wze and Wzb in Lactococcus lactis subsp. cremoris MG1363. In order to demonstrate the importance of ATP for the interactions among these proteins, native and fusion Wzb, Wzd and Wze proteins were expressed and purified from Escherichia coli cultures. The modulator protein, Wzd, binds with the phosphotyrosine kinase Wze, irrespective of its phosphorylation status. However, Wze attained a higher phosphorylation level after interacting with phosphorylated Wzd in the presence of 10 mM ATP. This highly phosphorylated Wze did not remain in close association with phosphorylated Wzd. The Wze tyrosine kinase protein of Lactobacillus rhamnosus thus carries out tyrosine phosphorylation of Wzd in addition to auto- and trans- phosphorylation of the kinase itself.