Drosophila melanogaster mounts a unique immune response to the Rhabdovirus sigma virus.

Rhabdoviruses are important pathogens of humans, livestock, and plants that are often vectored by insects. Rhabdovirus particles have a characteristic bullet shape with a lipid envelope and surface-exposed transmembrane glycoproteins. Sigma virus (SIGMAV) is a member of the Rhabdoviridae and is a naturally occurring disease agent of Drosophila melanogaster. The infection is maintained in Drosophila populations through vertical transmission via germ cells. We report here the nature of the Drosophila innate immune response to SIGMAV infection as revealed by quantitative reverse transcription-PCR analysis of differentially expressed genes identified by microarray analysis. We have also compared and contrasted the immune response of the host with respect to two nonenveloped viruses, Drosophila C virus (DCV) and Drosophila X virus (DXV). We determined that SIGMAV infection upregulates expression of the peptidoglycan receptor protein genes PGRP-SB1 and PGRP-SD and the antimicrobial peptide (AMP) genes Diptericin-A, Attacin-A, Attacin-B, Cecropin-A1, and Drosocin. SIGMAV infection did not induce PGRP-SA and the AMP genes Drosomycin-B, Metchnikowin, and Defensin that are upregulated in DCV and/or DXV infections. Expression levels of the Toll and Imd signaling cascade genes are not significantly altered by SIGMAV infection. These results highlight shared and unique aspects of the Drosophila immune response to the three viruses and may shed light on the nature of the interaction with the host and the evolution of these associations.

Evidence for multiplication of the leafhopper-borne maize yellow stripe virus in its vector using ELISA and dot-blot hybridization.

Maize yellow stripe virus (MYSV) has several features in common with tenuiviruses, but is transmitted by a leafhopper, Cicadulina chinai (Cicadellidae, Hemiptera), rather than planthoppers (Delphacidae, Hemiptera). Herein, MYSV was shown to be propagatively transmitted like tenuiviruses. MYSV RNA was not detected in leafhoppers by dot blot hybridization one day following a 2-day acquisition access period (AAP), but was detected in single or groups of leafhoppers 5-20 days post-acquisition. Likewise, capsid protein of MYSV was not detected by ELISA in single leafhoppers until the third day after the beginning of a 1- or 3-day AAP, but subsequently, mean absorbance values (405 nm) increased gradually, reaching their highest levels 8-14 days post-acquisition. The percentage of ELISA-positive leafhoppers also increased during the same period. Unlike most tenuiviruses, transovarial transmission of MYSV was not detected in 600 C. chinai nymphs that hatched from eggs laid by females that had acquired MYSV from diseased plants. The implications of our findings for MYSV classification are discussed.

Mutations in the potyvirus helper component protein: effects on interactions with virions and aphid stylets.

Mutations of K --> E in the highly conserved 'KITC' motif of the potyvirus helper component (HC) protein result in loss of HC function in aphid transmission, presumably because of inability to interact with virions, stylets or both. In this study we show that HC of potato virus C (PVC), a naturally occurring variant of potato virus Y (PVY) that has the K --> E mutation, lacks the ability to be retained in stylets, whereas PVY HC is retained. The K --> E mutation in either PVC or a site-directed mutant of tobacco etch virus (TEV) did not hinder binding to capsid protein, nor did deletion of the N-terminal 107 aa of TEV HC. An additional mutation, F -->, L at aa 10 of TEV HC, which renders HC non-functional but does not affect binding to capsid protein, is reported. Collectively, the results suggest that the N-terminal domain is required for interaction of HC with stylets rather than for binding to virions.

Association of the non-structural P3 viral protein with cylindrical inclusions in potyvirus-infected cells.

Antibodies raised against the 42K non-structural P3 protein encoded by the RNA genome of a potyvirus (tobacco vein mottling virus, TVMV) have been used with immunogold labelling procedures to determine the subcellular location of P3 in infected Nicotiana tabacum cells. P3-specific gold label was found almost exclusively associated with the cylindrical inclusions typically formed in the cytoplasm of potyvirus-infected cells by another non-structural viral protein, the cylindrical inclusion protein. The P3 antibodies reacted with inclusions in both the transverse (pinwheel-like) and longitudinal (bundle-like) orientations of these structures. Immunocytological examination of TVMV-infected protoplasts showed the association of P3 with cylindrical inclusions during the early stages of formation of these structures and suggests that the P3 may be involved in the replication of potyviral RNA.

Assembly and accumulation sites of maize mosaic virus in its planthopper vector.

The morphology, assembly, and accumulation sites of rhabdovirus particles in Peregrinus maidis planthoppers infected with a Hawaiian isolate of maize mosaic virus (MMV) were studied. These particles were usually bullet-shaped, but were sometimes bacilliform, and averaged 234 and 247 nm, respectively, in length and 60 nm in width. They were found in most acini of the principal and accessory salivary glands and in brain, nerve ganglia, leg muscle, foregut, midgut, trachea, epidermis, and fat and connective tissues. In most tissues MMV particles accumulated mainly within intracytoplasmic, dilated cisternae that were connected to the perinuclear space. However, in the salivary glands virus particles accumulated mainly in intercellular and extracellular spaces and were found in secretion vesicles. MMV particles appeared to bud from three types of membranes: (i) inner, and rarely outer, nuclear membranes of cells in most tissues examined; (ii) intracytoplasmic membranes, e.g., endoplasmic reticulum in salivary glands; and (iii) plasma membranes of salivary gland cells and nerve axons. The plasma membrane has not been reported previously as a budding site for plant rhabdoviruses, although it is known as a major assembly site for animal rhabdoviruses.