Production of succinoglycan polymer in Sinorhizobium meliloti is affected by SMb21506 and requires the N-terminal domain of ExoP.
The protein tyrosine kinase ExoP, consisting of an N-terminal periplasmic and a C-terminal cytoplasmic domain, is important for polymerization of the exopolysaccharide succinoglycan (EPS I) in Sinorhizobium meliloti. We analyzed the contribution of the ExoP paralogs ExoP2 and SMb21506 to the production of the high molecular weight (HMW) form of EPS I. ExoP2, though not contributing to EPS I or lipopolysaccharide biosynthesis, showed increased expression at high osmolarity and was expressed in Medicago sativa nodules, suggesting an involvement in the synthesis of an as-yet-unidentified polysaccharide. Furthermore, a mutation in SMb21506 affected the production of HMW EPS I, particularly in the absence of the C-terminal ExoP domain. High salinity induced the production of HMW EPS I by the wild type and mutants whereas high osmolarity had the opposite effect. It was shown that ExoP localizes at the inner membrane of S. meliloti cells. Tyrosine phosphorylation of the C-terminal domain was strongly increased by amino acid substitutions in the polysaccharide co-polymerase motif (formerly proline-rich motif) located in the N-terminal domain, suggesting that this phosphorylation could be modulated by conformational changes of the N-terminal domain. Moreover, deletion of a coiled-coil motif present in the N-terminal domain abolished phosphorylation and EPS I production and, consequently, the ability to nodulate M. sativa.
Jofré
Edgardo
E
Departamento de Ciencias Naturales, Facultad de Ciencias Exactas, Fisico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina. ejofre@exa.unrc.edu.ar
Becker
Anke
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D000455:Medicago sativa / Q000382:microbiology; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D055170:Plant Root Nodulation / Q000502:physiology; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis*; D017434:Protein Structure, Tertiary; D012329:RNA, Bacterial / Q000235:genetics; D016962:Sinorhizobium meliloti / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; C095765:ExoP protein, Rhizobium meliloti; D026901:Membrane Transport Proteins; D011135:Polysaccharides, Bacterial; D012329:RNA, Bacterial; C056779:succinoglycan