Characterization of a Listeria monocytogenes protein interfering with Rab5a.
Listeria monocytogenes (LM) phagocytic strategy implies recruitment and inhibition of Rab5a. Here, we identify a Listeria protein that binds to Rab5a and is responsible for Rab5a recruitment to phagosomes and impairment of the GDP/GTP exchange activity. This protein was identified as a glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Listeria (p40 protein, Lmo 2459). The p40 protein was found within the phagosomal membrane. Analysis of the sequence of LM p40 protein revealed two enzymatic domains: the nicotinamide adenine dinucleotide (NAD)-binding domain at the N-terminal and the C-terminal glycolytic domain. The putative ADP-ribosylating ability of this Listeria protein located in the N-terminal domain was examined and showed some similarities to the activity and Rab5a inhibition exerted by Pseudomonas aeruginosa ExoS onto endosome-endosome fusion. Listeria p40 caused Rab5a-specific ADP ribosylation and blocked Rab5a-exchange factor (Vps9) and GDI interaction and function, explaining the inhibition observed in Rab5a-mediated phagosome-endosome fusion. Meanwhile, ExoS impaired Rab5-early endosomal antigen 1 (EEA1) interaction and showed a wider Rab specificity. Listeria GAPDH might be the first intracellular gram-positive enzyme targeted to Rab proteins with ADP-ribosylating ability and a putative novel virulence factor.
Alvarez-Dominguez
Carmen
C
Servicio de Inmunología and Instituto de Formación e Investigación Marqués de Valdecilla (IFIMAV), Hospital Universitario "Marqués de Valdecilla", 39008 Santander, Spain. calvarez@humv.es
Madrazo-Toca
Fidel
F
Fernandez-Prieto
Lorena
L
Vandekerckhove
Joël
J
Pareja
Eduardo
E
Tobes
Raquel
R
Gomez-Lopez
Maria Teresa
MT
Del Cerro-Vadillo
Elida
E
Fresno
Manuel
M
Leyva-Cobián
Francisco
F
Carrasco-Marín
Eugenio
E
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002999:Clone Cells; D011992:Endosomes / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008089:Listeria monocytogenes / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D051379:Mice; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D010588:Phagosomes / Q000378:metabolism; D020696:rab5 GTP-Binding Proteins / Q000378:metabolism*
D001426:Bacterial Proteins; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D020696:rab5 GTP-Binding Proteins