Reciprocal secretion of proteins by the bacterial type III machines of plant and animal pathogens suggests universal recognition of mRNA targeting signals.

Bacterial pathogens of both animals and plants use type III secretion machines to inject virulence proteins into host cells. Although many components of the secretion machinery are conserved among different bacterial species, the substrates for their type III pathways are not. The Yersinia type III machinery recognizes some secretion substrates via a signal that is encoded within the first 15 codons of yop mRNA. These signals can be altered by frameshift mutations without affecting secretion of the encoded polypeptides, suggesting a mechanism whereby translation of yop mRNA is coupled to the translocation of newly synthesized polypeptide. We report that the type III machinery of Erwinia chrysanthemi cloned in Escherichia coli recognizes the secretion signals of yopE and yopQ. Pseudomonas syringae AvrB and AvrPto, two proteins exported by the recombinant Erwinia machine, can also be secreted by the Yersinia type III pathway. Mapping AvrPto sequences sufficient for the secretion of reporter fusions in Yersinia revealed the presence of an mRNA secretion signal. We propose that 11 conserved components of type III secretion machines may recognize signals that couple mRNA translation to polypeptide secretion.

[1]  H. Wolf‐Watz,et al.  Functional conservation of the secretion and translocation machinery for virulence proteins of yersiniae, salmonellae and shigellae. , 1995, The EMBO journal.

[2]  G. Cornelis,et al.  The Yersinia Yop virulon: a bacterial system for subverting eukaryotic cells , 1997, Molecular microbiology.

[3]  Dara W. Frank The exoenzyme S regulon of Pseudomonas aeruginosa , 1997, Molecular microbiology.

[4]  C. Hueck,et al.  Type III Protein Secretion Systems in Bacterial Pathogens of Animals and Plants , 1998, Microbiology and Molecular Biology Reviews.

[5]  A. Collmer,et al.  The type III (Hrp) secretion pathway of plant pathogenic bacteria: trafficking harpins, Avr proteins, and death , 1997, Journal of bacteriology.

[6]  G. Cornelis,et al.  Restriction of DNA in Yersinia enterocolitica detected by recipient ability for a derepressed R factor from Escherichia coli. , 1975, Journal of general microbiology.

[7]  R. Rosqvist,et al.  Intracellular targeting of exoenzyme S of Pseudomonas aeruginosa via type III‐dependent translocation induces phagocytosis resistance, cytotoxicity and disruption of actin microfilaments , 1997, Molecular microbiology.

[8]  J. Miller,et al.  Effects of surrounding sequence on the suppression of nonsense codons. , 1983, Journal of molecular biology.

[9]  O. Schneewind,et al.  Two independent type III secretion mechanisms for YopE in Yersinia enterocolitica , 1997, Molecular microbiology.

[10]  J. Galán,et al.  The invasion-associated type-III protein secretion system in Salmonella--a review. , 1997, Gene.

[11]  G. Cornelis,et al.  Customized secretion chaperones in pathogenic bacteria , 1996, Molecular microbiology.

[12]  O. Schneewind,et al.  Yersinia enterocolitica Type III Secretion , 1999, The Journal of Biological Chemistry.

[13]  S. He,et al.  Hrp pilus: an hrp-dependent bacterial surface appendage produced by Pseudomonas syringae pv. tomato DC3000. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[14]  G. Stormo,et al.  Translational initiation in prokaryotes. , 1981, Annual review of microbiology.

[15]  G. Cornelis,et al.  Analysis of virC, an operon involved in the secretion of Yop proteins by Yersinia enterocolitica , 1991, Journal of bacteriology.

[16]  A. Pühler,et al.  A new family of RSF1010-derived expression and lac-fusion broad-host-range vectors for gram-negative bacteria. , 1990, Gene.

[17]  O. Schneewind,et al.  Type III machines of pathogenic yersiniae secrete virulence factors into the extracellular milieu , 1999, Molecular microbiology.

[18]  G. Cornelis,et al.  Individual chaperones required for Yop secretion by Yersinia. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[19]  D. Fouts,et al.  A cloned Erwinia chrysanthemi Hrp (type III protein secretion) system functions in Escherichia coli to deliver Pseudomonas syringae Avr signals to plant cells and to secrete Avr proteins in culture. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[20]  E. Skrzypek,et al.  Regulation by Ca2+ in the Yersinia low‐Ca2+ response , 1993, Molecular microbiology.

[21]  E. Skrzypek,et al.  Yops of Yersinia spp. pathogenic for humans , 1993, Infection and immunity.

[22]  S. He,et al.  Pseudomonas syringae pv. syringae harpinPss: A protein that is secreted via the hrp pathway and elicits the hypersensitive response in plants , 1993, Cell.

[23]  O. Schneewind,et al.  Targeting of Yersinia Yop proteins into the cytosol of HeLa cells: one‐step translocation of YopE across bacterial and eukaryotic membranes is dependent on SycE chaperone , 1998, Molecular microbiology.

[24]  I. Lambermont,et al.  Identification of the YopE and YopH domains required for secretion and internalization into the cytosol of macrophages, using the cyaA gene fusion approach. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[25]  O. Schneewind,et al.  A mRNA signal for the type III secretion of Yop proteins by Yersinia enterocolitica. , 1997, Science.

[26]  M. Zuker On finding all suboptimal foldings of an RNA molecule. , 1989, Science.

[27]  K. Hardie,et al.  The C‐terminal domain of the secretin PulD contains the binding site for its cognate chaperone, PulS, and confers PulS dependence on pIVf1 function , 1997, Molecular microbiology.

[28]  O. Schneewind,et al.  Type III secretion machines and the pathogenesis of enteric infections caused by Yersinia and Salmonella spp. , 1999, Immunological reviews.

[29]  O. Schneewind,et al.  Yersinia enterocolitica type III secretion: an mRNA signal that couples translation and secretion of YopQ , 1999, Molecular microbiology.

[30]  D. Hanahan Studies on transformation of Escherichia coli with plasmids. , 1983, Journal of molecular biology.

[31]  R M Macnab,et al.  Genetics and biogenesis of bacterial flagella. , 1992, Annual review of genetics.

[32]  O. Schneewind,et al.  Type III machines of Gram-negative pathogens: injecting virulence factors into host cells and more. , 1999, Current opinion in microbiology.

[33]  G. Cornelis,et al.  The Virulence Plasmid of Yersinia, an Antihost Genome , 1998, Microbiology and Molecular Biology Reviews.

[34]  P. Wattiau,et al.  SycE, a chaperone‐like protein of Yersinia enterocolitica involved in the secretion of YopE , 1993, Molecular microbiology.

[35]  J. Kaper EPEC delivers the goods. , 1998, Trends in microbiology.

[36]  J. Tommassen,et al.  The outer membrane component, YscC, of the Yop secretion machinery of Yersinia enterocolitica forms a ring‐shaped multimeric complex , 1997, Molecular microbiology.