Natively unfolded nucleic acid binding P8 domain of SeMV polyprotein 2a affects the novel ATPase activity of the preceding P10 domain

Nair, Smita; Savithri, H.S.

doi:10.1016/j.febslet.2009.12.003

Cited by 23 publications

(26 citation statements)

References 18 publications

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“…In a broader sense, the interaction of P1 with the genome-bound VPg and P10 was considered as a cue of an active transport of viral RNA complex facilitated by P1 acting as a MP. The energy needed for that process was suspected to come from the hydrolysis of ATP by SeMV P10, as P1 itself did not show any ATPase activity [ 99 , 100 ]. When expressed in E. coli , SeMV P1 appeared to be in large soluble aggregates, characteristic of several MPs [ 98 ].…”

Section: P1 In Viral Movement and Suppression Of Rna Silencingmentioning

confidence: 99%

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Overview on Sobemoviruses and a Proposal for the Creation of the Family Sobemoviridae

Sõmera

Sarmiento

Truve

2015

Viruses

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The genus Sobemovirus, unassigned to any family, consists of viruses with single-stranded plus-oriented single-component RNA genomes and small icosahedral particles. Currently, 14 species within the genus have been recognized by the International Committee on Taxonomy of Viruses (ICTV) but several new species are to be recognized in the near future. Sobemovirus genomes are compact with a conserved structure of open reading frames and with short untranslated regions. Several sobemoviruses are important pathogens. Moreover, over the last decade sobemoviruses have become important model systems to study plant virus evolution. In the current review we give an overview of the structure and expression of sobemovirus genomes, processing and functions of individual proteins, particle structure, pathology and phylogenesis of sobemoviruses as well as of satellite RNAs present together with these viruses. Based on a phylogenetic analysis we propose that a new family Sobemoviridae should be recognized including the genera Sobemovirus and Polemovirus. Finally, we outline the future perspectives and needs for the research focusing on sobemoviruses.

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Section: P1 In Viral Movement and Suppression Of Rna Silencingmentioning

confidence: 99%

“…The C-terminal part of P2a is not conserved. It was demonstrated that SeMV P2a C-terminus contains an RNA-binding domain (P10) and a novel ATPase domain (P8) [ 100 ]. The C-terminal part of the polyprotein P2ab consists of the motifs characteristic for an RdRp.…”

Section: Proteolytic Processing Of Polyproteinmentioning

confidence: 99%

Overview on Sobemoviruses and a Proposal for the Creation of the Family Sobemoviridae

Sõmera

Sarmiento

Truve

2015

Viruses

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“…It is not clear whether it has a functional role, or if it is only the byproduct of proteolytic cleavage. Recent studies using purified ORF2a encoded C-terminal fragments from SeMV, demonstrated ATPase and nucleic acid binding activity for the corresponding recombinant proteins (Nair and Savithri, 2010a). Such properties are characteristic, for example, for plant virus-encoded helicases.…”

Section: Introductionmentioning

confidence: 99%

Expression and characterisation of the ryegrass mottle virus non-structural proteins

Baļķe¹,

Resevica²,

Skrastiņa³

et al. 2010

Proceedings of the Latvian Academy of Sciences. Section B. Natural, Exact, and Applied Sciences.

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Expression and characterisation of the ryegrass mottle virus non-structural proteins The Ryegrass mottle virus (RGMoV) single-stranded RNA genome is organised into four open reading frames (ORF) which encode several proteins: ORF1 encodes protein P1, ORF2a contains the membrane-associated 3C-like serine protease, genome-linked protein VPg and a P16 protein gene. ORF2b encodes replicase RdRP and the only structural protein, coat protein, is synthesised from ORF3. To obtain the non-structural proteins in preparative quantities and to characterise them, the corresponding RGMoV gene cDNAs were cloned in pET- and pColdI-derived expression vectors and overexpressed in several E. coli host cells. For protease and RdRP, the best expression system containing pColdI vector and E. coli WK6 strain was determined. VPg and P16 proteins were obtained from the pET- or pACYC- vectors and E. coli BL21 (DE3) host cells and purified using Ni-Sepharose affinity chromatography. Attempts to crystallize VPg and P16 were unsuccessful, possibly due to non-structured amino acid sequences in both protein structures. Methods based on bioinformatic analysis indicated that the entire VPg domain and the C-terminal part of the P16 contain unstructured amino acid stretches, which possibly prevented the formation of crystals.

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“…For example, it was shown that the protease-VPg (Δ70 Pro-VPg) but not the protease (Δ70 Pro) alone is active [15]. Similarly, the ATPase activity of p10 domain was stimulated by the p8 domain present at its C-terminus [14]. Further, VPg-RdRp is the predominant intermediate of 2ab processing in E.coli [11].…”

Section: Introductionmentioning

confidence: 99%

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

2012

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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.

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Natively unfolded nucleic acid binding P8 domain of SeMV polyprotein 2a affects the novel ATPase activity of the preceding P10 domain

Cited by 23 publications

References 18 publications

Overview on Sobemoviruses and a Proposal for the Creation of the Family Sobemoviridae

Overview on Sobemoviruses and a Proposal for the Creation of the Family Sobemoviridae

Expression and characterisation of the ryegrass mottle virus non-structural proteins

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

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