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Comparative analysis of the full genome sequence of European bat lyssavirus type 1 and type 2 with other lyssaviruses and evidence for a conserved transcription termination and polyadenylation motif in the G–L 3' non-translated region

¹ Rabies & Wildlife Zoonoses Group, Veterinary Laboratories Agency (VLA, Weybridge), WHO Collaborating Centre for the Characterisation of Rabies and Rabies-Related Viruses, New Haw, Addlestone, Surrey KT15 3NB, UK
² Institute for Epidemiology, WHO Collaborating Centre for Rabies Surveillance and Research, OIE Reference Laboratory for Rabies, Friedrich Loeffler Institute - Federal Research Institute for Animal Health, Seestrasse 55, D-16868 Wusterhausen, Germany
³ Max-von-Pettenkofer Institute and Gene Center, Feodor-Lynen-Str. 25, D-81377 Munich, Germany
⁴ Unité Stratégies Antivirales, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France

Correspondence
A. R. Fooks
t.fooks{at}vla.defra.gsi.gov.uk

We report the first full-length genomic sequences for European bat lyssavirus type-1 (EBLV-1) and type-2 (EBLV-2). The EBLV-1 genomic sequence was derived from a virus isolated from a serotine bat in Hamburg, Germany, in 1968 and the EBLV-2 sequence was derived from a virus isolate from a human case of rabies that occurred in Scotland in 2002. A long-distance PCR strategy was used to amplify the open reading frames (ORFs), followed by standard and modified RACE (rapid amplification of cDNA ends) techniques to amplify the 3' and 5' ends. The lengths of each complete viral genome for EBLV-1 and EBLV-2 were 11 966 and 11 930 base pairs, respectively, and follow the standard rhabdovirus genome organization of five viral proteins. Comparison with other lyssavirus sequences demonstrates variation in degrees of homology, with the genomic termini showing a high degree of complementarity. The nucleoprotein was the most conserved, both intra- and intergenotypically, followed by the polymerase (L), matrix and glyco- proteins, with the phosphoprotein being the most variable. In addition, we have shown that the two EBLVs utilize a conserved transcription termination and polyadenylation (TTP) motif, approximately 50 nt upstream of the L gene start codon. All available lyssavirus sequences to date, with the exception of Pasteur virus (PV) and PV-derived isolates, use the second TTP site. This observation may explain differences in pathogenicity between lyssavirus strains, dependent on the length of the untranslated region, which might affect transcriptional activity and RNA stability.

Tables showing the primers and the accession numbers for the sequences used in this study are available as supplementary material in JGV Online.

The GenBank/EMBL/DDBJ accession numbers for the full genome sequences of EBLV-1 and -2 are EF157976 and EF157977.

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