title: Mutations of arginine residues within the 146-KKRRK-150 motif of the ActA protein of Listeria monocytogenes abolish intracellular motility by interfering with the recruitment of the Arp2/3 complex.
abstract: The recruitment of actin to the surface of intracellular Listeria monocytogenes and subsequent tail formation is dependent on the expression of the bacterial surface protein ActA. Of the different functional domains of ActA identified thus far, the N-terminal region is absolutely required for actin filament recruitment and intracellular motility. Mutational analysis of this domain which abolished actin recruitment by intracellular Listeria monocytogenes identified two arginine residues within the 146-KKRRK-150 motif that are essential for its activity. More specifically, recruitment of the Arp2/3 complex to the bacterial surface, as assessed by immunofluorescence staining with antibodies raised against the p21-Arc protein, was not obtained in these mutants. Consistently, treatment of infected cells with latrunculin B, which abrogated actin filament formation, did not affect association of ActA with p21-Arc at the bacterial surface. Thus, the initial recruitment of the Arp2/3 complex to the bacterial surface is independent of, and precedes, actin polymerisation. Our data suggest that binding of the Arp2/3 complex is mediated by specific interactions dependent on arginine residues within the 146-KKRRK-150 motif present in ActA.
authors.affiliation: Department of Cell Biology, Gesellschaft für Biotechnologische Forschung, Mascheroder Weg 1, D-38124 Braunschweig, Germany. spi@biobase.de
authors.forename: S
authors.initials: S
authors.lastname: Pistor
authors.forename: L
authors.initials: L
authors.lastname: Gröbe
authors.forename: A S
authors.initials: AS
authors.lastname: Sechi
authors.forename: E
authors.initials: E
authors.lastname: Domann
authors.forename: B
authors.initials: B
authors.lastname: Gerstel
authors.forename: L M
authors.initials: LM
authors.lastname: Machesky
authors.forename: T
authors.initials: T
authors.lastname: Chakraborty
authors.forename: J
authors.initials: J
authors.lastname: Wehland
mesh_terms: D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002876:Chromosomes, Bacterial; D003598:Cytoskeletal Proteins; D005798:Genes, Bacterial; D007424:Intracellular Fluid / Q000378:metabolism; D008089:Listeria monocytogenes / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D017384:Sequence Deletion; D013844:Thiazoles / Q000378:metabolism; D053778:Thiazolidines
chemical_list: D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; D001426:Bacterial Proteins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D013844:Thiazoles; D053778:Thiazolidines; C072980:actA protein, Listeria monocytogenes; D001120:Arginine; C037068:latrunculin B