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Although previous studies of Bunyaviridae mRNA termination sites have been successful in mapping the termination position, the methods employed were limited to such that the position could only be determined within 4 or 5 nucleotides (Bouloy et al., 1984). Furthermore, non-viral sequences such as poly-A tail, at the 3' end of mRNAs would not be detected by these mapping techniques. Because of these shortcomings, we attempted to clone and sequence intact 3' ends of Hantaan virus S genome-derived antigenomic RNAs using the crossover sequencing method. We report here the analysis of the 5' terminus of S genomic RNA and the 3' terminus of S-specific mRNAs using the crossover sequencing technique. We found that S genomic RNA contains only two trinucleotide repeats at its 5' end and the 3' ends of mRNAs are truncated and are not polyadenylated. The hantavirus genus of family Bunyaviridae is comprised of several serologically distinct virus types; Hantaan virus is the prototype of the genus hantavirus which is the etiologic agent of hemorrhagic fever with renal syndrome (HFRS). Hantaviruses are enveloped negative sense RNA viruses which contain three genomic segments; large (L), medium (M), and small (S). All three segments of many different strains of hantaviruses have been cloned and sequenced All the previous reports showed that there are three trinucleotide repeats at both the 3' and the 5' ends of all hantavirus genomic Nucleotide sequence analysis of the L-segments revealed that L-segments contain approximately 6,500 nucleotides which has a single open reading frame to code for approximately 2,150 amino acids (Schmaljohn, 1990; Antic et al. , 1991b; Stohwasser et al., 1991). The M segments of hantaviruses contain approximately 3,700 nucleotides which also has a single open reading frame which The product of M segment is cleaved into G1 and G2 glycoproteins with approximate molecular masses of 68 and 55 kDa respectively. The S segments of hantaviruses contain approximately 1,700 nucleotides which code for nucleocapsid protein of approximately 430 amino acids (Schmaljohn et al. We have recently reported that 5' ends of Hantaan virus mRNA contains host cell derived cap at the 5' end (Garcin et al., 1995). This is consistent with previous studies that Bunyavirus, Phlebovirus and Uukuvirus genera which initiate mRNA transcription via a cellular cap-stealing mechanism, a process known to occur with influenza virus systems (Plotch et al., 1981). The RNase T2 and S1 nuclease mapping techniques (Eshita et al., 1985; Bouloy et …

experimental and molecular medicine/experimental and molecular medicine

The S-genomic RNA of Hantaan virus contains only two trinucleotide repeats at its 5' end and the 3' end of S-specific mRNA is truncated and lacks poly-A tail