Genetic characterization of K13965, a strain of Oak Vale virus from Western Australia

Abstract

K13965, an uncharacterized virus, was isolated in 1993 from Anopheles annulipes mosquitoes collected in the Kimberley region of northern Western Australia. Here, we report its genomic sequence, identify it as a rhabdovirus, and characterize its phylogenetic relationships. The genome comprises a P' (C) and SH protein similar to the recently characterized Tupaia and Durham viruses, and shows overlap between G and L genes. Comparison of K13965 genome sequence to other rhabdoviruses identified K13965 as a strain of the unclassified Australian Oak Vale rhabdovirus, whose complete genome sequence we also determined. Phylogenetic analysis of N and L sequences indicated genetic relationship to a recently proposed Sandjima virus clade, although the Oak Vale virus sequences form a branch separate from the African members of that group.

Figure 1

Map of Australia by states and territories showing the geographic locations where OVRV K13965 and CSIRO-1342 were isolated. K13965 was isolated from Anopheles annulipes s.l. collected 3 km north of Kununurra. OVRV CSIRO-1342 was isolated from Culex edwardsi collected at Peachester. Another isolation of OVRV has been reported from Aedes vigilax collected at Darwin. ACT, Australian Capital Territory; NSW, New South Wales; NT, Northern Territory; QLD, Queensland; SA, South Australia; TAS, Tasmania; VIC, Victoria, WA, Western Australia.

Figure 2

K13965 genome organization. (A) Schematic organization of the K13965 genome. The bar represents the 11,120 nucleotide (nt) antigenome. The eight open arrows indicate the position of the open reading frames (ORFs) for putative N, P, P′, P″, M, SH, G, and L proteins. (B) Transcription initiation, intergenic and transcription termination/polyadenylation sequences. The start and stop codon for each gene is in bold, the intergenic region is underlined. (C) Complementarity between the 3′ leader and the 5′ trailer sequences. Bases conserved among rhabdoviruses known to infect vertebrates are shown in bold. Start and stop codons are underlined.

Figure 3

Phylogenetic relationships of K13965 and CSIRO-1342 with representative rhabdoviruses derived from partial L (156 aa; A), N (325 aa; B) and full L amino acid sequence (970 aa; C). The geographic origin (*Africa, # Australia) is indicated for K13965, CSIRO-1342 and members of the Sandjimba and Almpiwar groups. K13965 is indicated by a circled arrowhead. Analysis was performed using a Bayesian method applying a WAG model of amino acid replacement with a gamma distribution of rate variation among sites for 10 million generations (with a 25% burn-in). Support for each node is provided by BPP (Bayesian posterior probability) (>0.8). Branch lengths are drawn to scale and the trees were mid-point rooted. Amino acid sequences used were: ABLV, Australian bat lyssavirus (L:NP_478343; N:AAD01267); ALMV, Almpiwar virus (L:AY854645); ARAV, Aravan virus (L:ABV03822; N:Q6X1D8); ARV, Adelaide River virus (L:AF234534; N:AAC54627); BBOV, Bimbo virus (L:GU816016); BEFV, bovine ephemeral fever virus (L:NP_065409; N:NP_065398); BGNV, Bangoran virus (L:GU816010); BRMV, Berrimah virus (L:AAZ43265); BTKV, Boteke virus (L:GU816014); BYSMV, barley yellow striate mosaic virus (L:ACT21686); CFRV, China fish rhabdovirus (L:AAX86686); CHPV, Chandipura virus (L:P13179; N:P11211); CHVV9824, Charleville virus (L:AY854644); CHVV9847, Charleville virus (L:AY854672); COCV, Cocal virus (L:ACB47438; N:ACB47434); CPV, coastal plains virus (L:ADG86364; N:ADG86356); DAffSV, Drosophila affinis sigma virus 10 (L:GQ410980); DobsSV, Drosophila obscura sigma virus 10A (N:GQ410979); DURV, Durham virus (L:ADB88761; N:ADB88758); DUVV, Duvenhage virus (L:ABZ81216; N:Q66453); EBLV1, european bat lyssavirus 1 (L:ABZ81181; N:AAX62875); EBLV2, european bat lyssavirus 2 (L:ABZ81191; N:YP_001285393); FLAV, Flanders virus (L:AAN73288; N:AAN73283); FUKV, Fukuoka virus (L:AAZ43279); GARV, Garba virus (L:GU816018); HDOOV, Humpty doo virus (L:AAZ43271); HIRRV, Hirame rhabdovirus (L:NP_919035; N:NP_919030); IHNV, infectious hematopoietic necrosis virus (L:CAA52076; N:NP042676); IRKV, Irkut virus (L:ABV03823; N:Q5VKP6); ISFV, Isfahan virus (L:Q5K2K3; N:Q5K2K7); KAMV, Kamese virus (L:GU816011); KCV, Kern Canyon virus (N:ABE69215); KEUV, Keuraliba virus (L:GU816021); KHUV, Khujand virus (L:ABV03824; N:Q6X1D4); KIMV, Kimberley virus (L:AAZ43266); KOLV, Kolongo virus (L:GU816020; N:ABE69214); KOTV, Kotonkan virus (L:AAZ43267; N:ABE69213); LBV, Lagos bat virus (L:ABZ81171; N:ABF56214); LDV, Le Dantec virus (L:AAZ43278); LYMoV, lettuce yellow mottle virus (L:YP_002308376; N:YP_00208371); LNYV, lettuce necrotic yellows virus (L:YP_425092; N:CAG34083); MEBV, mount Elgon bat virus (N:ABE69217); MFSV, maize fine streak virus (L:YP_052849; N:YP_052843); MMV, maize mosaic virus (L:YP_052855; N:YP_052850); MOKV, Mokola virus (L:ABZ81211; N:YP_142350); MOSV, Mossuril virus (L:GU816012); MOUV, Moussa virus (L:ACZ81402; N:ACZ81403); NASV, Nasoule virus (L:GU816017); NCMV, northern cereal mosaic virus (L:NP_597914; N:NP_057954); NGAV, Ngaingan virus (L:YP_003518294; N:YP_003518280); NKOV, Nkolbisson virus (L:GU816022); OBOV, Obodhiang virus (N:ABE69212); OFV, orchid fleck virus (L:YP_001294929; N:BAH97109); OITAV, Oita rhabdovirus (N:BAD13431); OUAV, Ouango virus (L:GU816015); PCRV, Parry Creek virus (L:AAZ43275); PERV, Perinet virus (L:AAZ43280); PFRV, Pike fry rhabdovirus (L:ACP28002; N:ACP27998); PIRYV, Piry virus (N:P26037); PORV, Porton virus (L:GU816013); PYDV, potato yellow dwarf virus (N:ABW35154); RABV, Rabies virus (L:Q66T60; N:ACN15666); RBUV, Rochambeau virus (N:ABE69218); RYSV, rice yellow stunt virus (L:NP_620502; N:NP_620496); SCRV, Siniperca chuatsi rhabdovirus (L:YP_802942; N:YP_802937); SCV, strawberry crinkle virus (L:AAP03645); SFRV, Starry flounder rhabdovirus (L:AY450644); SHRV, snakehead rhabdovirus (L:NP_050585; N:NP_050580); SIGMAV, Drosophila melanogaster sigma virus (N:ACV67011); SIGMAVHAP23, Drosophila melanogaster sigma virus HAP23 (L:GQ375258; N:GQ375258); SIGMAVHAP30, Drosophila melanogaster sigma virus AP30 (L:YP_003126913; N:YP_003126908); SJAV, Sandjimba virus (L:GU816019; N:ABE69216); STRV, sea trout rhabdovirus (N:AAL35756); SVCV, spring viraemia of carp virus (L:Q91DR9; N:ABW24033); SYNV, Sonchus yellow net virus (L:NP_042286; N:NP_042281); TaVCV, Taro vein chlorosis virus (L:YP_224083; N:YP_224078); TIBV, Tibrogargan virus (L:ADG86355; N:ADG86347); TUPV, Tupaia virus (L:YP_238534; N:YP_238528); VHSV, viral hemorrhagic septicemia virus (L:CAB40833; N:P24378); VSAV, vesicular stomatitis Alagoas virus (L:ACB47443; N:ACB47439); VSIV, vesicular stomatitis Indiana virus (L:NP_041716; N:P11212); VSNJV, vesicular stomatitis New Jersey virus (L:P16379; N:P04881); WCBV, West Caucasian bat virus (L:ABV03821; N:Q5VKP2); WOGV, Wongabel virus (L:AAZ43276; N:YP_002333271).