.::) 1992 Oxford University Press

Nucleic Acids Research, Vol. 20, No. 12 3249

Complete nucleotide sequence of a neuropathogenic variant of Friend murine leukemia virus PVC-211 Mary P.Remington, Paul M.Hoffman, Sandra K.Ruscettil and Michiaki Masuda1 * Retrovirus Research Center, Veterans Administration Medical Center, Baltimore, MD 21218 and 1Laboratory of Molecular Oncology, National Cancer Institute, Frederick, MD 21702-1201, USA Submitted May 14, 1992

GenBank accession

PVC-2 1 1 murine leukemia virus (MuLV) is a replication ecotropic MuLV which was derived by passaging Friend MuLV through Fischer rats (1). It causes a rapidly progressive neurological disease in susceptible rats and mice (1, 2), but unlike the wild type Friend MuLV, PVC-211 MuLV does not induce acute erythroleukemia in mice (2, 3). The characteristic pathological changes in the nervous sytems of the animals with the neurological disease are spongiform vacuolation, perivascular astrogliosis and neuronal degeneration in the brain stem, cerebellum and spinal cord (2). Interestingly, viral replication is detected primarily in brain capillary endothelial cells but not in glial cells or neurons (2). We have isolated an infectious molecular clone (3d) of PVC-211 MuLV to study the molecular determinants of the disease specificity of this virus (3). PVC-211 clone 3d containing the entire viral genome was ligated into pUC19 at the unique EcoRI site within the pol gene (3) (base 2850 of the given sequence). Subgenomic DNA fragments were prepared and cloned into either pUC 18, pUCl9 or pGEM-7Zf(+) for sequence determination. The nucleotide sequence was determined by the dideoxy chain termination method on double-stranded plasmid DNA templates by using either Sequenase 2.0 (U.S. Biochemical, Cleveland, Ohio) and 32P-dATP or the Taq dye primer cycle sequencing system and an automated DNA sequencer (373A; Applied Biosystems, Inc., Foster City, California). The nucleotide sequence of the LTR, which is responsible for the attenuated leukemogenicity of PVC-2 11 MuLV, and that of the env gene, which is necessary for the neuropathogenicity of the virus, were reported previously (3). Here, we report the complete nucleotide sequence of the PVC-2 11 MuLV clone 3d. The level of homology between PVC-2 11 and other strains of Friend MuLV, 57 (4), FB29 (5) and K-1 (6, 7), is also presented in the Table. That PVC-211 MuLV shares the highest degree of homology with the strain K- 1 is compatible with the fact that both of these strains were derived from a Friend virus stock given by Dr C.Friend to the Cancer Institue in Tokyo, Japan (1, 6). competent

ACKNOWLEDGEMENTS We thank Dr Hiroshi Amanuma for the unpublished

To whom correspondence should be addressed

1. 2. 3. 4.

5. 6. 7. 8.

Kai,K. and Furuta,T. (1984) J. Virol. 50, 970-973. Hoffman,P.M. et al. (1992) Lab. Invest. in press. Masuda,M. et al. (1992) J. Virol. 66, 2798-2806. European Molecular Biology Laboratory (EMBL) Data Library, accession number X02794. Perryman,S. et al. (1991) Nucleic Acids Res. 24, 6950. Obata,M. et al. (1984) Virology 136, 435-438. Amanuma,H. Personal communication. Devereux,J. et al. (1984) Nucleic Acids Res. 12, 387-395.

Table. Sequence homology between PVC-211 clone 3d and other strains of Friend MuLVa

U3 R U5 5' leaderb gag po/ env

strain 57

strain FB29

strain K- 1

81.7 91.2 98.7 95.4 97.8 (96.5)d 97.9 (98.4) 97.2 (96.0)

87.4 97.1 98.7 95.4 98.3 (97.0) 98.4 (98.7) 97.7 (96.3)

97.9 98.5 98.7

N.A." N.A. N.A. 99.3 (98.5)

aThe nucleotide sequences or amino acid sequences were aligned by the GAP program of the GCG sequence analysis software package (8), and the percent identity was calculated by counting a gap as a mismatch. bThe 5' leader sequence extends from the 3' end of U5 to the ATG gag start codon. CN.A. = data not available. dNumbers in parentheses indicate the percent identity of deduced amino acid sequence. (Linear) nAPPLOT of: Pvc200.Seg ck: 3389. 0 to: 8220 April 7. 0992 t9 14. 2000 1000 3000 7000 4000 soo0 6000

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of the LTR of Friend MuLV strain K-1. We also thank Denise Rubens (Laboratory of Mathematical Biology, National Cancer Institute) for facilitating the automated sequencing. *

REFERENCES

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