A non-toxic pokeweed antiviral protein mutant inhibits pathogen infection via a novel salicylic acid-independent pathway.
Pokeweed antiviral protein (PAP), a ribosome-inactivating protein isolated from Phytolacca americana, is characterized by its ability to depurinate the sarcin/ricin (S/R) loop of the large rRNA of prokaryotic and eukaryotic ribosomes. In this study, we present evidence that PAP is associated with ribosomes and depurinates tobacco ribosomes in vivo by removing more than one adenine and a guanine. A mutant of pokeweed antiviral protein, PAPn, which has a single amino acid substitution (G75D), did not bind ribosomes efficiently, indicating that Gly-75 in the N-terminal domain is critical for the binding of PAP to ribosomes. PAPn did not depurinate ribosomes and was non-toxic when expressed in transgenic tobacco plants. Unlike wild-type PAP and a C-terminal deletion mutant, transgenic plants expressing PAPn did not have elevated levels of acidic pathogenesis-related (PR) proteins. PAPn, like other forms of PAP, did not trigger production of salicylic acid (SA) in transgenic plants. Expression of the basic PR proteins, the wound-inducible protein kinase and protease inhibitor II, was induced in PAPn-expressing transgenic plants and these plants were resistant to viral and fungal infection. These results demonstrate that PAPn activates a particular SA-independent, stress-associated signal transduction pathway and confers pathogen resistance in the absence of ribosome binding, rRNA depurination and acidic PR protein production.
Zoubenko
O
O
Department of Plant Pathology, Cook College, Rutgers University, New Brunswick, NJ 08903, USA.
Hudak
K
K
Tumer
N E
NE
D001483:Base Sequence; D015870:Gene Expression; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics; D009154:Mutation; D009699:N-Glycosyl Hydrolases; D010935:Plant Diseases / Q000235:genetics* / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011485:Protein Binding; D011687:Purines / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1; D012270:Ribosomes / Q000378:metabolism; D020156:Salicylic Acid / Q000378:metabolism*; D015398:Signal Transduction; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology; D015533:Transcriptional Activation
D010940:Plant Proteins; D011687:Purines; D012335:RNA, Ribosomal; D054789:Ribosome Inactivating Proteins, Type 1; C053376:pathogenesis-related proteins, plant; D020928:Mitogen-Activated Protein Kinases; C113423:WIPK protein, Nicotiana tabacum; D009699:N-Glycosyl Hydrolases; C024275:pokeweed antiviral protein; D020156:Salicylic Acid; C030985:purine