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The Spike Protein of Potato Yellow Dwarf Virus and Its Functional Role in the Infection of Insect Vector Cells

Biological Institute, University of Stuttgart, Ulmerstrasse 227, D-7000 Stuttgart 60, F.R.G.

Treatment of particles of potato yellow dwarf virus, serotype SYDV, with non-ionic detergents solubilized two of the five structural proteins, G and M₁. Since only these proteins were also recovered in lipid vesicles prepared either by a detergent dialysis technique or after organic extraction of purified virus using phosphatidylcholine in n-hexane, it is suggested that M₂ protein is not membrane-associated but rather belongs to the core. The SYDV spike protein G was purified by isoelectric focusing during which it appeared to be heterogeneous in charge with most of the protein banding at pH 4.8. The same heterogeneity was observed after two-dimensional protein gel electrophoresis, where SYDV G protein was found largely at pH 4.8, whereas that of the other PYDV serotype, CYDV, was detected largely at pH 4.3. Antibodies raised against purified SYDV G protein reacted specifically with the G protein. When they were present during the inoculation step, G-specific antibodies were able to neutralize the infectivity of SYDV for insect vector cell monolayers, which suggests that G protein has a functional role in the inoculation process. Isolated SYDV G protein or SYDV envelope vesicles containing G protein inhibited the infection of vector cells by SYDV; however, only with SYDV envelope vesicles was the inhibition concentration-dependent. This indicated competition between virions and the vesicles during the inoculation process, possibly for recognition or attachment sites.

Keywords: PYDV, glycoprotein isolation, infection, lipid vesicles

Received 25 February 1986; accepted 21 August 1986.