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Bovine respiratory syncytial virus nucleocapsid protein expressed in insect cells specifically interacts with the phosphoprotein and the M2 protein.

The coding region of the gene for the nucleocapsid (N) protein of bovine respiratory syncytial virus has been inserted into the genome of Autographa californica nuclear polyhedrosis virus (AcNPV) using transfer vector pVL 1393. Infection of Spodoptera frugiperda cells with recombinant virus resulted in the synthesis of high levels of N protein. This protein was indistinguishable from the authentic bovine respiratory syncytial virus (BRSV) N protein by SDS-gel electrophoresis and immunoprecipitation with anti-BRSV serum. [35S]methionine-labeled N protein synthesized in S. frugiperda cells was used in a protein-blotting protein overlay assay for its interactions with BRSV-infected cell proteins. The N protein synthesized in S. frugiperda cells was found to specifically interact with the phosphoprotein (P) and M2 protein of BRSV but only with the P protein of human RSV. Our results suggest that the recombinant N protein produced in S. frugiperda cells could be used to determine the domains on N protein required for binding to P and M2 proteins.

Animals↗

Efficient whitefly transmission of African cassava mosaic geminivirus requires sequences from both genomic components.

Clones of two subgroup III geminiviruses, the common strain of tomato golden mosaic virus (csTGMV) and African cassava mosaic virus originating from Kenya (ACMV-K), were shown to be non-transmissible by whitefiles. Lack of transmissibility of cloned ACMV-K was investigated by exchanging genomic components with a whitefly-transmissible ACMV isolate from Nigeria (ACMV-NOg). Neither pseudorecombinant was transmissible, indicating that defects in both genomic components contributed to the lack of transmissibility. Analysis of the acquisition of the pseudorecombinats by Bemisia tabaci indicated that accumulation of virus within the insect was DNA B dependent. Return of virus to plants was determined by DNA A, although the coat protein was essential for acquisition. Repeated passaging of both the wild strain of ACMV-NOg and the cloned virus led to loss of insect transmissibility of the wild isolate but not the cloned virus. Products encoded on both genomic components are required for transmission of bipartite geminiviruses by B. tabaci.

Animals↗

The 5' untranslated region of Rhopalosiphum padi virus contains an internal ribosome entry site which functions efficiently in mammalian, plant, and insect translation systems.

Rhopalosiphum padi virus (RhPV) is one of several picorna-like viruses that infect insects; sequence analysis has revealed distinct differences between these agents and mammalian picornaviruses. RhPV has a single-stranded positive-sense RNA genome of about 10 kb; unlike the genomes of Picornaviridae, however, this genome contains two long open reading frames (ORFs). ORF1 encodes the virus nonstructural proteins, while the downstream ORF, ORF2, specifies the structural proteins. Both ORFs are preceded by long untranslated regions (UTRs). The intergenic UTR is known to contain an internal ribosome entry site (IRES) which directs non-AUG-initiated translation of ORF2. We have examined the 5' UTR of RhPV for IRES activity by translating synthetic dicistronic mRNAs containing this sequence in a variety of systems. We now report that the 5' UTR contains an element which directs internal initiation of protein synthesis from an AUG codon in mammalian, plant, and Drosophila in vitro translation systems. In contrast, the encephalomyocarditis virus IRES functions only in the mammalian system. The RhPV 5' IRES element has features in common with picornavirus IRES elements, in that no coding sequence is required for IRES function, but also with cellular IRES elements, as deletion analysis indicates that this IRES element does not have sharply defined boundaries.

5' Untranslated Regions↗

Persistence of invertebrate iridescent virus 6 in tropical artificial aquatic environments. Brief report.

The rate of loss of activity of invertebrate iridescent virus 6 (IIV-6, family Iridoviridae) was determined in two artificial aquatic habitats in southern Mexico, using a sensitive insect bioassay technique. IIV-6 placed in trays of water in direct sunlight suffered rapid loss of activity (99.99% reduction) over a period of 36 h, during which water temperatures fluctuated between 24 and 41 degrees C. No significant deactivation occurred during the hours of darkness. In contrast, IIV-6 placed in trays of water in the shade lost 97% of original activity over a 60 h period, during which water temperatures fluctuated from 24 to 31 degrees C. Longitudinal analysis involving mixed effects models of time (shade) and cumulative exposure to ultraviolet radiation (sunlight) indicated that the rate of deactivation was best described by third order polynomial equations in both cases. We conclude that the likelihood of transmission of IIVs in aquatic habitats will be mediated by the intensity of UV radiation and water temperature.

Animals↗