title: Complete gene sequence of a human Puumala hantavirus isolate, Puumala Umeå/hu: sequence comparison and characterisation of encoded gene products.
abstract: Puumala virus is a member of the hantavirus genus in the Bunyaviridae family, and the major causative agent of haemorrhagic fever with renal syndrome in Europe. This study was conducted with a human Puumala virus isolate (PUUV Umeå/hu), and contains the determination of the first complete PUUV sequence from a human source. When the relationship to other Puumala viruses was analysed, a possible RNA segment exchange between two local strains of PUUV was noticed. Furthermore, the coding regions of PUUV Umeå/hu S- and M-segments were cloned, and a large set of gene products were expressed in mammalian cells. In addition, postulated N- and O-linked glycosylation sites in the two envelope proteins (Gn and Gc) were investigated individually by site-directed mutagenesis followed by gel-shift analysis. Our data demonstrate that N-linked glycosylation occurs at three sites in Gn (N142, N357 and N409), and at one site in Gc (N937). Also, one possible O-glycosylation site was identified in Gc (T985). We conclude that the diversity between different Puumala virus isolates is high, and consequently characterization of local PUUV isolates is important for clinical diagnostic work. Finally, the obtained results concerning the encoded gene products are of great importance for the design of new vaccines.
authors.affiliation: Department of Medical Countermeasures, Division of NBC Defence, Swedish Defence Research Agency, SE-901 82 Umeå, Sweden.
authors.forename: Patrik
authors.initials: P
authors.lastname: Johansson
authors.forename: Marie
authors.initials: M
authors.lastname: Olsson
authors.forename: Lena
authors.initials: L
authors.lastname: Lindgren
authors.forename: Clas
authors.initials: C
authors.lastname: Ahlm
authors.forename: Fredrik
authors.initials: F
authors.lastname: Elgh
authors.forename: Anna
authors.initials: A
authors.lastname: Holmström
authors.forename: Göran
authors.initials: G
authors.lastname: Bucht
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary; D024202:Electrophoretic Mobility Shift Assay; D015870:Gene Expression; D005814:Genes, Viral; D016679:Genome, Viral; D006031:Glycosylation; D006480:Hemorrhagic Fever with Renal Syndrome / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010802:Phylogeny; D011499:Protein Processing, Post-Translational; D029262:Puumala virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D012367:RNA, Viral; D014759:Viral Envelope Proteins