title: Posttranslocation chaperone PrsA2 regulates the maturation and secretion of Listeria monocytogenes proprotein virulence factors.
abstract: PrsA2 is a conserved posttranslocation chaperone and a peptidyl prolyl cis-trans isomerase (PPIase) that contributes to the virulence of the Gram-positive intracellular pathogen Listeria monocytogenes. One of the phenotypes associated with a prsA2 mutant is decreased activity of the broad-range phospholipase C (PC-PLC). PC-PLC is made as a proenzyme whose maturation is mediated by a metalloprotease (Mpl). The proforms of PC-PLC and Mpl accumulate at the membrane-cell wall interface until a decrease in pH triggers their maturation and rapid secretion into the host cell. In this study, we examined the mechanism by which PrsA2 regulates the activity of PC-PLC. We observed that in the absence of PrsA2, the proenzymes are secreted at physiological pH and do not mature upon a decrease in pH. The sensitivity of the prsA2 mutant to cell wall hydrolases was modified. However, no apparent changes in cell wall porosity were detected. Interestingly, synthesis of PC-PLC in the absence of its propeptide lead to the secretion of a fully active enzyme in the cytosol of host cells independent of PrsA2, indicating that neither the propeptide of PC-PLC nor PrsA2 is required for native folding of the catalytic domain, although both influence secretion of the enzyme. Taken together, these results suggest that PrsA2 regulates compartmentalization of Mpl and PC-PLC, possibly by influencing cell wall properties and interacting with the PC-PLC propeptide. Moreover, the ability of these proproteins to respond to a decrease in pH during intracellular growth depends on their localization at the membrane-cell wall interface.
authors: Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853-6401, USA.
authors: Brian M
authors: BM
authors: Forster
authors: Jason
authors: J
authors: Zemansky
authors: Daniel A
authors: DA
authors: Portnoy
authors: Hélène
authors: H
authors: Marquis
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008089:Listeria monocytogenes / Q000201:enzymology* / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000378:metabolism*; D019696:Peptidylprolyl Isomerase / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D010738:Type C Phospholipases / Q000378:metabolism*; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D011498:Protein Precursors; D037521:Virulence Factors; D010738:Type C Phospholipases; C105598:phosphatidylcholine-specific phospholipase C; D008666:Metalloendopeptidases; C080367:mpl protein, Listeria monocytogenes; D019696:Peptidylprolyl Isomerase; C541924:PrsA2 protein, Listeria monocytogenes