12940993

pubmed:Citation

5

The Listeria monocytogenes surface protein ActA mediates actin-based motility by interacting with a number of host cytoskeletal components, including Ena/VASP family proteins, which in turn interact with actin and the actin-binding protein profilin. We employed a bidirectional genetic approach to study Ena/VASP's contribution to L. monocytogenes movement and pathogenesis. We generated an ActA allelic series within the defined Ena/VASP-binding sites and introduced the resulting mutant L. monocytogenes into cell lines expressing different Ena/VASP derivatives. Our findings indicate that Ena/VASP proteins contribute to the persistence of both speed and directionality of L. monocytogenes movement. In the absence of the Ena/VASP proline-rich central domain, speed consistency decreased by sixfold. In addition, the Ena/VASP F-actin-binding region increased directionality of bacterial movement by fourfold. We further show that both regions of Ena/VASP enhanced L. monocytogenes cell-to-cell spread to a similar degree, although the Ena/VASP F-actin-binding region did so in an ActA-independent manner. Surprisingly, our ActA allelic series enabled us to uncouple L. monocytogenes speed from directionality although both were controlled by Ena/VASP proteins. Lastly, we showed the pathogenic relevance of these findings by the observation that L. monocytogenes lacking ActA Ena/VASP-binding sites were up to 400-fold less virulent during an adaptive immune response.

eng

IM

MEDLINE

0950-382X

Print

NLM

1361-75

pubmed-meshheading:12940993-Actins, pubmed-meshheading:12940993-Animals, pubmed-meshheading:12940993-Bacterial Proteins, pubmed-meshheading:12940993-Binding Sites, pubmed-meshheading:12940993-Carrier Proteins, pubmed-meshheading:12940993-Cell Adhesion Molecules, pubmed-meshheading:12940993-Cell Line, pubmed-meshheading:12940993-Cytoskeletal Proteins, pubmed-meshheading:12940993-Genes, Bacterial, pubmed-meshheading:12940993-Listeria monocytogenes, pubmed-meshheading:12940993-Listeriosis, pubmed-meshheading:12940993-Macrophages, pubmed-meshheading:12940993-Membrane Proteins, pubmed-meshheading:12940993-Mice, pubmed-meshheading:12940993-Mice, Inbred C57BL, pubmed-meshheading:12940993-Microfilament Proteins, pubmed-meshheading:12940993-Movement, pubmed-meshheading:12940993-Mutation, Missense, pubmed-meshheading:12940993-Phosphoproteins, pubmed-meshheading:12940993-Point Mutation, pubmed-meshheading:12940993-Protein Binding, pubmed-meshheading:12940993-Protein Structure, Secondary, pubmed-meshheading:12940993-Protein Structure, Tertiary, pubmed-meshheading:12940993-Rats, pubmed-meshheading:12940993-Sequence Deletion, pubmed-meshheading:12940993-Virulence

Ena/VASP proteins contribute to Listeria monocytogenes pathogenesis by controlling temporal and spatial persistence of bacterial actin-based motility.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't