Cis- and Trans-acting Elements in Cowpea Mosaic Virus RNA Replication

Bokhoven, Hans van; Gall, Olivier Le; Kasteel, D.; Verver, J.; Wellink, J.; Kammen, A. B. Van

doi:10.1006/viro.1993.1387

Cited by 61 publications

(53 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…3c). Previous studies have shown that the 110 kDa protein is stable in protoplasts (van Bokhoven et al, 1993) and data presented below indicated that 110-YFP-Zm7hvr was also a stable protein. Protoplasts inoculated with pMON-110-YFP-Zm7hvr and CPMV showed accumulation of 110-YFP-Zm7hvr in tubules (Fig.…”

Section: Mp Molecules Interact Within the Tubulementioning

confidence: 59%

See 1 more Smart Citation

Studies on the origin and structure of tubules made by the movement protein of Cowpea mosaic virus

Pouwels¹,

Velden²,

Willemse³

et al. 2004

Journal of General Virology

View full text Add to dashboard Cite

Cowpea mosaic virus (CPMV) moves from cell to cell by transporting virus particles via tubules formed through plasmodesmata by the movement protein (MP). On the surface of protoplasts, a fusion between the MP and the green fluorescent protein forms similar tubules and peripheral punctate spots. Here it was shown by time-lapse microscopy that tubules can grow out from a subset of these peripheral punctate spots, which are dynamic structures that seem anchored to the plasma membrane. Fluorescence resonance energy transfer experiments showed that MP subunits interacted within the tubule, where they were virtually immobile, confirming that tubules consist of a highly organized MP multimer. Fluorescence recovery after photobleaching experiments with protoplasts, transiently expressing fluorescent plasma membrane-associated proteins of different sizes, indicated that tubules made by CPMV MP do not interact directly with the surrounding plasma membrane. These experiments indicated an indirect interaction between the tubule and the surrounding plasma membrane, possibly via a host plasma membrane protein. INTRODUCTIONFor successful systemic infection, a plant virus must spread throughout the plant, a process that starts with transport from the initially infected cell to neighbouring uninfected cells (cell-to-cell movement) and is followed by transport through the phloem into roots and young developing leaves (systemic movement). Since plant viruses cannot pass through the rigid cell wall, they have evolved ways of exploiting plasmodesmata, the naturally occurring transport channels present between plant cells (Haywood et al., 2002). Normally, only small molecules are able to pass through plasmodesmata, but plant viruses encode one or more proteins, the so-called movement proteins (MPs), that modify the structure of plasmodesmata in such a way that viral transport is enabled. So far, two basic principles for cell-to-cell movement of plant viruses have been described: tubule-guided movement of virus particles and movement as ribonucleoprotein complexes (Lazarowitz & Beachy, 1999).Cowpea mosaic virus (CPMV), a positive-stranded, bipartite RNA virus belonging to the family Comoviridae, moves from cell to cell by transporting virus particles using tubular structures, which connect the infected cell to the neighbouring uninfected cell (Pouwels et al., 2002a). Immunogold labelling has shown that the CPMV MP is present in these tubular structures (van Lent et al., 1990). On the surface of CPMV-infected protoplasts, similar tubular structures are formed, which protrude up to 20 mm into the culture medium, are tightly surrounded by the plasma membrane and have the same ultrastructure as tubules in plant tissue (van Lent et al., 1991). Remarkably, tubules are also formed on protoplasts transiently expressing MP , showing that MP is the only viral protein required for tubule formation. So far, protoplasts have proved extremely useful as a model system for studying targeting and assembly of both wt and mutant MPs (Bertens et al., 20...

show abstract

Section: Mp Molecules Interact Within the Tubulementioning

confidence: 59%

“…Protoplasts were isolated from cowpea (Vigna unguiculata L. 'California Blackeye') leaves and transfected as described previously (van Bokhoven et al, 1993). For cytoskeleton inhibitor studies, protoplasts were divided into two aliquots: one was left untreated and the other was treated with 20 mM latrunculin B or 10 mM oryzalin.…”

Section: Methodsmentioning

confidence: 99%

Studies on the origin and structure of tubules made by the movement protein of Cowpea mosaic virus

Pouwels¹,

Velden²,

Willemse³

et al. 2004

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…Data in which nonsense and frameshift mutations prevented RNA replication even in the presence of helper genomes have also been reported for mouse hepatitis virus (de Groot et al 1992), clover yellow mosaic virus {White et al 1992), cowpea mosaic virus (van Bokhoven et al 1993}, and turnip yellow mosaic virus (Weiland and Dreher 1993}, although in none of these studies were possible effects of mutations on RNA sequence and structure excluded. Nevertheless, several other cytoplasmic positive-strand RNA viruses that can, like poliovirus, use the same RNA molecules both as a mRNA and as a template for negative-strand synthesis, might also employ a cis requirement for translation.…”

Section: Why Should An Rna Virus Require Translation In Cis?mentioning

confidence: 93%

Coupling between genome translation and replication in an RNA virus.

Novak¹,

Kirkegaard²

1994

Genes Dev.

181

175

View full text Add to dashboard Cite

The replication of poliovirus RNA genomes containing amber mutations was studied to test whether viral proteins provided in trans could rescue the replication of an RNA genome that could not be completely translated itself. Mutants containing amber codons at different positions in the genome displayed vastly different abilities to be rescued by wild-type proteins provided by a helper genome. Amber-suppressing cell lines were used to ensure that the defects in the amber mutants arose from their failure to be translated, not from defects in RNA sequence or structure. An internal region of the poliovirus genome was identified whose translation is required in cis; failure to translate this region was shown to inhibit RNA replication. This coupling between translation and RNA replication could provide a late proofreading mechanism that enables poliovirus, and possibly many other RNA viruses, to prevent the replication of defective genomes.

show abstract

“…Whereas the AUG at 115 is out of frame with that at 512 and has no known function (Wellink et al, 1993b), the AUG at position 161 is in-frame with AUG 512 and is functional as an initiation codon (Holness et al, 1989). Either deleting AUG 161 or disrupting its frame relationship with AUG 512 effectively eliminates RNA-2 replication (Holness et al, 1989;van Bokhoven et al, 1993). The need to preserve the frame relationship between AUG 161 and 512 to retain the replication ability of RNA-2-based constructs complicates the construction of vectors (Sainsbury et al, 2008b).…”

mentioning

confidence: 99%

Extremely High-Level and Rapid Transient Protein Production in Plants without the Use of Viral Replication

2008

View full text Add to dashboard Cite

Plant-based overexpression of heterologous proteins has attracted much interest and development in recent years. To date, the most efficient vectors have been based on RNA virus-derived replicons. A system based on a disabled version of cowpea mosaic virus RNA-2 has been developed, which overcomes limitations on insert size and introduces biocontainment. This system involves positioning a gene of interest between the 5# leader sequence and 3# untranslated region (UTR) of RNA-2, thereby emulating a presumably stable mRNA for efficient translation. Thus far, the sequence of the 5# UTR has been preserved to maintain the ability of the modified RNA-2 to be replicated by RNA-1. However, high-level expression may be achieved in the absence of RNA-1-derived replication functions using Agrobacterium-mediated transient transformation. To investigate those features of the 5# UTR necessary for efficient expression, we have addressed the role of two AUG codons found within the 5# leader sequence upstream of the main initiation start site. Deletion of an in-frame start codon upstream of the main translation initiation site led to a massive increase in foreign protein accumulation. By 6 d postinfiltration, a number of unrelated proteins, including a full-size IgG and a self-assembling virus-like particle, were expressed to .10% and 20% of total extractable protein, respectively. Thus, this system provides an ideal vehicle for high-level expression that does not rely on viral replication of transcripts.

show abstract

Cis- and Trans-acting Elements in Cowpea Mosaic Virus RNA Replication

Cited by 61 publications

References 0 publications

Studies on the origin and structure of tubules made by the movement protein of Cowpea mosaic virus

Studies on the origin and structure of tubules made by the movement protein of Cowpea mosaic virus

Coupling between genome translation and replication in an RNA virus.

Extremely High-Level and Rapid Transient Protein Production in Plants without the Use of Viral Replication

Contact Info

Product

Resources

About