The complete nucleotide sequence of the S RNA of tomato spotted wilt virus (TSWV) was determined. The RNA is 2916 nucleotides long and has an ambisense coding strategy. The sequence contains two open reading frames (ORFs), one in the viral sense which encodes a protein with a predicted Mr of 52.4K and one in the viral complementary sense which encodes the viral nucleocapsid protein of Mr 28.8K. Both proteins are expressed by translation of two subgenomic RNA species that possibly terminate at a long stable hairpin structure, located at the intergenic region. The structure of this RNA segment resembles that of the arthropod-borne phleboviruses (family Bunyaviridae). The absence of significant sequence homology between TSWV and bunyaviruses infecting animals suggests that TSWV should be considered as a representative of a new genus within the Bunyaviridae.