Identification, Purification, and Serological Detection of the Major Noncapsid Protein of Rice Grassy Stunt Virus

Miranda, G. J.; Koganezawa, H.

doi:10.1094/phyto-85-1530

“…Falk et al (39) identified a nonstructural protein of maize stripe tenuivirus that accumulated in maize but not in the leafhopper vector. Similar results were recently reported for the related rice grassy stunt tenuivirus in rice and its leafhopper host (103). The function of the nonstructural protein is unknown.…”

Section: Circulative Propagative Transmissionsupporting

confidence: 87%

Mechanisms of Arthropod Transmission of Plant and Animal Viruses

Gray

¹

,

Banerjee

²

1999

Microbiol Mol Biol Rev

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SUMMARY A majority of the plant-infecting viruses and many of the animal-infecting viruses are dependent upon arthropod vectors for transmission between hosts and/or as alternative hosts. The viruses have evolved specific associations with their vectors, and we are beginning to understand the underlying mechanisms that regulate the virus transmission process. A majority of plant viruses are carried on the cuticle lining of a vector’s mouthparts or foregut. This initially appeared to be simple mechanical contamination, but it is now known to be a biologically complex interaction between specific virus proteins and as yet unidentified vector cuticle-associated compounds. Numerous other plant viruses and the majority of animal viruses are carried within the body of the vector. These viruses have evolved specific mechanisms to enable them to be transported through multiple tissues and to evade vector defenses. In response, vector species have evolved so that not all individuals within a species are susceptible to virus infection or can serve as a competent vector. Not only are the virus components of the transmission process being identified, but also the genetic and physiological components of the vectors which determine their ability to be used successfully by the virus are being elucidated. The mechanisms of arthropod-virus associations are many and complex, but common themes are beginning to emerge which may allow the development of novel strategies to ultimately control epidemics caused by arthropod-borne viruses.

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“…5A) showed that fraction 10 contained the majority of the ribulose bisphosphate carboxylase/oxygenase complex (Rubisco), an oligomeric complex of ϳ560 kDa composed of multiple copies of both a 53-kDa large subunit and a 14.9-kDa small subunit. These 20 samples were also run on four other SDS-PAGE (12.5% polyacrylamide) gels, which were separately subjected to Western blot analysis with antisera against p2 (5), p5 (5), p6 (14), and RNP particles (9) to identify fractions containing the p2, p5, p6, and N proteins. Fractions 3, 4, and 5 contained the most p5, and fractions 2, 3, and 4 contained the most p4, whereas N was found primarily in fraction 20, the bottom fraction, presumably in RNP particles (data not shown).…”

Section: Vol 77 2003 Notes 771mentioning

confidence: 99%

Detection of rice grassy stunt tenuivirus nonstructural proteins p2, p5 and p6 from infected rice plants and from viruliferous brown planthoppers

Chomchan

¹

,

Miranda

²

,

Shirako

³

2002

Archives of Virology

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We investigated the interaction of Rice grassy stunt tenuivirus (RGSV) nonstructural protein p5, a protein of 22 kDa encoded on vRNA 5, with all 12 RGSV proteins by using a GAL4 transcription activator-based yeast two-hybrid system. The p5 protein interacted only with itself and not with any other viral protein; the interacting domains were localized within the N-terminal 96 amino acids of p5. The p5-p5 interaction was reproduced in an Sos recruitment-mediated yeast two-hybrid system as well in by far-Western blots. Native p5 protein extracted from RGSV-infected rice tissue was detected in a large complex with a molecular mass of approximately 260 kDa composed of 12 molecules of p5 or a p5 oligomer with an unidentified host factor(s).The genome of Rice grassy stunt tenuivirus (RGSV), a member in the genus Tenuivirus (12) and an important rice pathogen in East, Southeast, and South Asian countries (8), consists of six ambisense RNA segments containing a total of 12 open reading frames (ORFs) (13,16,17). RGSV replicates and is transmitted by a brown planthopper (Nilaparvata lugens Stal) (9). So far, little has been known about the functions of the individual RGSV proteins, except for the 339-kDa RNA-dependent RNA polymerase (RdRp) encoded on the complementary strand of RNA 1 (cRNA 1) and the 36-kDa nucleocapsid protein (N) encoded on cRNA 5 ( Fig. 1), both of which are found along with genomic and possibly complementary RNAs as thin filamentous ribonucleoprotein (RNP) particles. Recently, we showed that the 22-kDa p5 protein encoded on the virus genomic strand of RNA 5 (vRNA 5) accumulates in large amounts in both RGSV-infected rice leaves and viruliferous brown planthoppers, while the 23-kDa p2 protein encoded on vRNA 2 and the 21-kDa p6 protein encoded on vRNA 6 were preferentially expressed in infected rice leaf tissues rather than in viruliferous insects (5). We hypothesized that p5 has an essential role in virus infection in both the plant and insect hosts (5).In this study, we investigated possible interactions between the p5 protein and the other RGSV proteins by using a GAL4 transcription activator-based yeast two-hybrid system (4, 6). We found that p5 interacts with itself through domains located in the N-terminal half of the protein, but does not interact with any other RGSV protein with this yeast two-hybrid system. The p5-p5 interaction was confirmed by an Sos recruitment-mediated yeast two-hybrid system (1, 2) as well as by far-Western blots, indicating that the strength of the p5-p5 interaction is significant and suggesting that the interaction is biologically significant as well. p5-p5 interaction in the N-terminal region detected by a GAL4 transcription activator-based yeast two-hybrid system.Interactions between the p5 protein and all 12 RGSV proteins were examined by using a yeast two-hybrid system based on the GAL4 transcription activator (4, 6) (MatchMaker 2; Clontech). In this system, the yeast GAL4 transcription activator has been separated into two functional domains: (i) the DNA binding act...

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“…5A) showed that fraction 10 contained the majority of the ribulose bisphosphate carboxylase/oxygenase complex (Rubisco), an oligomeric complex of ϳ560 kDa composed of multiple copies of both a 53-kDa large subunit and a 14.9-kDa small subunit. These 20 samples were also run on four other SDS-PAGE (12.5% polyacrylamide) gels, which were separately subjected to Western blot analysis with antisera against p2 (5), p5 (5), p6 (14), and RNP particles (9) to identify fractions containing the p2, p5, p6, and N proteins. Fractions 3, 4, and 5 contained the most p5, and fractions 2, 3, and 4 contained the most p4, whereas N was found primarily in fraction 20, the bottom fraction, presumably in RNP particles (data not shown).…”

mentioning

confidence: 99%

Rice Grassy Stunt Tenuivirus Nonstructural Protein p5 Interacts with Itself To Form Oligomeric Complexes In Vitro and In Vivo

Chomchan¹,

Li

²

,

Shirako

³

2003

J Virol

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We investigated the interaction of Rice grassy stunt tenuivirus (RGSV) nonstructural protein p5, a protein of 22 kDa encoded on vRNA 5, with all 12 RGSV proteins by using a GAL4 transcription activator-based yeast two-hybrid system. The p5 protein interacted only with itself and not with any other viral protein; the interacting domains were localized within the N-terminal 96 amino acids of p5. The p5-p5 interaction was reproduced in an Sos recruitment-mediated yeast two-hybrid system as well in by far-Western blots. Native p5 protein extracted from RGSV-infected rice tissue was detected in a large complex with a molecular mass of approximately 260 kDa composed of 12 molecules of p5 or a p5 oligomer with an unidentified host factor(s).

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Identification, Purification, and Serological Detection of the Major Noncapsid Protein of Rice Grassy Stunt Virus

Cited by 11 publications

References 0 publications

Mechanisms of Arthropod Transmission of Plant and Animal Viruses

Mechanisms of Arthropod Transmission of Plant and Animal Viruses

Detection of rice grassy stunt tenuivirus nonstructural proteins p2, p5 and p6 from infected rice plants and from viruliferous brown planthoppers

Rice Grassy Stunt Tenuivirus Nonstructural Protein p5 Interacts with Itself To Form Oligomeric Complexes In Vitro and In Vivo

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