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.

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