Location of Repository

Sesbania Mosaic Virus (SeMV) Infectious Clone: Possible Mechanism of 3′ and 5′ End Repair and Role of Polyprotein Processing in Viral Replication

By Kunduri Govind, Kristiina Mäkinen and Handanahal S. Savithri

Abstract

Sesbania mosaic virus (SeMV) is a positive stranded RNA virus belonging to the genus Sobemovirus. Construction of an infectious clone is an essential step for deciphering the virus gene functions in vivo. Using Agrobacterium based transient expression system we show that SeMV icDNA is infectious on Sesbania grandiflora and Cyamopsis tetragonoloba plants. The efficiency of icDNA infection was found to be significantly high on Cyamopsis plants when compared to that on Sesbania grandiflora. The coat protein could be detected within 6 days post infiltration in the infiltrated leaves. Different species of viral RNA (double stranded and single stranded genomic and subgenomic RNA) could be detected upon northern analysis, suggesting that complete replication had taken place. Based on the analysis of the sequences at the genomic termini of progeny RNA from SeMV icDNA infiltrated leaves and those of its 3′ and 5′ terminal deletion mutants, we propose a possible mechanism for 3′ and 5′ end repair in vivo. Mutation of the cleavage sites in the polyproteins encoded by ORF 2 resulted in complete loss of infection by the icDNA, suggesting the importance of correct polyprotein processing at all the four cleavage sites for viral replication. Complementation analysis suggested that ORF 2 gene products can act in trans. However, the trans acting ability of ORF 2 gene products was abolished upon deletion of the N-terminal hydrophobic domain of polyprotein 2a and 2ab, suggesting that these products necessarily function at the replication site, where they are anchored to membranes

Topics: Research Article
Publisher: Public Library of Science
OAI identifier: oai:pubmedcentral.nih.gov:3280281
Provided by: PubMed Central
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://www.pubmedcentral.nih.g... (external link)
  • Suggested articles

    Preview

    Citations

    1. (2007). A novel cleavage site within the potato leafroll virus P1 polyprotein.
    2. (2000). Agrobacterium transient expression system as a tool for the isolation of disease resistance genes: application to the Rx2 locus in potato.
    3. (2003). An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus.
    4. (2007). Biological systems of the host cell involved in Agrobacterium infection.
    5. (2000). Characterization of VPg and the polyprotein processing of cocksfoot mottle virus (genus Sobemovirus).
    6. (2009). Cis-acting RNA elements in human and animal plus-strand RNA viruses.
    7. (1990). Complete nucleotide sequence of infectious Coxsackievirus B3 cDNA: two initial 59 uridine residues are regained during plus-strand RNA synthesis.
    8. (2001). Complete nucleotide sequence of Sesbania mosaic virus: a new virus species of the genus Sobemovirus.
    9. (2008). Construction of infectious cDNA clones for RNA viruses: Turnip crinkle virus.
    10. (2008). Construction of infectious clones for RNA viruses:
    11. (1998). Expression of the rice yellow mottle virus P1 protein in vitro and in vivo and its involvement in virus spread.
    12. (2010). How RNA viruses maintain their genome integrity.
    13. (1998). Identification of viral genes required for cell-to-cell movement of southern bean mosaic virus.
    14. (2004). In planta engineering of viral RNA replicons: efficient assembly by recombination of DNA modules delivered by Agrobacterium.
    15. (2011). Interaction of Sesbania mosaic virus movement protein with the coat protein–implications for viral spread.
    16. (2011). Interaction of Sesbania mosaic virus movement protein with VPg and P10: implication to specificity of genome recognition.
    17. (1974). Large plasmid in Agrobacterium tumefaciens essential for crown gallinducing ability.
    18. (1995). Luteovirus gene expression.
    19. (1992). Modified binary plant transformation vectors with the wild-type gene encoding NPTII.
    20. (2010). Natively unfolded nucleic acid binding P8 domain of SeMV polyprotein 2a affects the novel ATPase activity of the preceding P10 domain.
    21. (2005). Natively unfolded’’ VPg is essential for Sesbania mosaic virus serine protease activity.
    22. (2006). P1 protein of Cocksfoot mottle virus is indispensable for the systemic spread of the virus.
    23. (2009). pEAQ: versatile expression vectors for easy and quick transient expression of heterologous proteins in plants.
    24. (2004). Polyprotein processing: cis and trans proteolytic activities of Sesbania mosaic virus serine protease.
    25. (2010). Primer-independent initiation of RNA synthesis by SeMV recombinant RNA-dependent RNA polymerase.
    26. (1982). Purification and partial characterization of sesbania mosaic virus.
    27. (2009). Renilla luciferase-based quantitation of Potato virus A infection initiated with Agrobacterium infiltration of N. benthamiana leaves.
    28. (2005). Replication-independent expression of genome components and capsid protein of brome mosaic virus in planta: a functional role for viral replicase in RNA packaging.
    29. (2001). Silencing on the spot. Induction and suppression of RNA silencing in the Agrobacterium-mediated transient expression system.
    30. (1994). Sitedirected mutagenesis of double-stranded DNA by the polymerase chain reaction.
    31. (2011). Sobemovirus RNA linked to VPg over a threonine residue.
    32. (2008). Stacking interactions of W271 and H275 of SeMV serine protease with W43 of natively unfolded VPg confer catalytic activity to protease.
    33. (1995). Synthesis of an infectious full-length cDNA clone of rice yellow mottle virus and mutagenesis of the coat protein.
    34. (2008). T-DNA binary vectors and systems.
    35. (1991). The 59-terminal nucleotides of hepatitis A virus RNA, but not poliovirus RNA, are required for infectivity.
    36. (2010). The rice yellow mottle virus P1 protein exhibits dual functions to suppress and activate gene silencing.
    37. (2007). Truve E

    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.