A focal adhesion factor directly linking intracellularly motile Listeria monocytogenes and Listeria ivanovii to the actin-based cytoskeleton of mammalian cells.
The surface-bound ActA polypeptide of the intracellular bacterial pathogen Listeria monocytogenes is the sole listerial factor needed for recruitment of host actin filaments by intracellularly motile bacteria. Here we report that following Listeria infection the host vasodilator-stimulated phosphoprotein (VASP), a microfilament- and focal adhesion-associated substrate of both the cAMP- and cGMP-dependent protein kinases, accumulates on the surface of intracytoplasmic bacteria prior to the detection of F-actin 'clouds'. VASP remains associated with the surface of highly motile bacteria, where it is polarly located, juxtaposed between one extremity of the bacterial surface and the front of the actin comet tail. Since actin filament polymerization occurs only at the very front of the tail, VASP exhibits properties of a host protein required to promote actin polymerization. Purified VASP binds directly to the ActA polypeptide in vitro. A ligand-overlay blot using purified radiolabelled VASP enabled us to identify the ActA homologue of the related intracellular motile pathogen, Listeria ivanovii, as a protein with a molecular mass of approximately 150 kDa. VASP also associates with actin filaments recruited by another intracellularly motile bacterial pathogen, Shigella flexneri. Hence, by the simple expedient of expressing surface-bound attractor molecules, bacterial pathogens effectively harness cytoskeletal components to achieve intracellular movement.
Chakraborty
T
T
Institut für Medizinische Mikrobiologie, Giessen.
Ebel
F
F
Domann
E
E
Niebuhr
K
K
Gerstel
B
B
Pistor
S
S
Temm-Grove
C J
CJ
Jockusch
B M
BM
Reinhard
M
M
Walter
U
U
D000199:Actins / Q000096:biosynthesis / Q000502:physiology*; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002460:Cell Line; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D003599:Cytoskeleton / Q000469:parasitology / Q000502:physiology*; D017931:DNA Primers; D005347:Fibroblasts / Q000469:parasitology / Q000502:physiology / Q000648:ultrastructure; D017353:Gene Deletion; D017125:Genes, Protozoan; D006801:Humans; D008087:Listeria / Q000502:physiology*; D008089:Listeria monocytogenes / Q000235:genetics / Q000502:physiology*; D008322:Mammals; D008840:Microfilament Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012867:Skin / Q000469:parasitology / Q000648:ultrastructure; D012879:Skin Physiological Phenomena; D000098662:Vasodilator-Stimulated Phosphoprotein
D000199:Actins; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D008840:Microfilament Proteins; D010750:Phosphoproteins; D000098662:Vasodilator-Stimulated Phosphoprotein; D017868:Cyclic AMP-Dependent Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases