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Rhizobia are symbiotic soil bacteria able to intracellularly colonize legume nodule cells and form nitrogen-fixing symbiosomes therein. How the plant cell cytoskeleton reorganizes in response to rhizobium colonization has remained poorly understood especially because of the lack of an  in vitro  infection assay. Here, we report on the use of the heterologous HeLa cell model to experimentally tackle this question. We observed that the model rhizobium  Sinorhizobium meliloti , and other rhizobia as well, were able to trigger a major reorganization of actin cytoskeleton of cultured HeLa cells  in vitro . Cell deformation was associated with an inhibition of the three major small RhoGTPases Cdc42, RhoA and Rac1. Bacterial entry, cytoskeleton rearrangements and modulation of RhoGTPase activity required an intact  S. meliloti  biosynthetic pathway for queuosine, a hypermodifed nucleoside regulating protein translation through tRNA, and possibly mRNA, modification. We showed that an intact bacterial queuosine biosynthetic pathway was also required for effective nitrogen-fixing symbiosis of  S. meliloti  with its host plant  Medicago truncatula , thus indicating that one or several key symbiotic functions of  S. meliloti  are under queuosine control. We discuss whether the symbiotic defect of  que  mutants may originate, at least in part, from an altered capacity to modify plant cell actin cytoskeleton.

Queuosine Biosynthesis Is Required for Sinorhizobium meliloti-Induced Cytoskeletal Modifications on HeLa Cells and Symbiosis with Medicago truncatula