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Journal of Virology, May 2001, p. 4332-4342, Vol. 75, No. 9
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.9.4332-4342.2001

L1 Interaction Domains of Papillomavirus L2 Necessary for Viral Genome Encapsidation

Martin M. Okun,1 Patricia M. Day,1 Heather L. Greenstone,1 Frank P. Booy,2 Douglas R. Lowy,1 John T. Schiller,1 and Richard B. S. Roden1,3,*

Laboratory of Cellular Oncology, Division of Basic Sciences, National Cancer Institute, Bethesda, Maryland 208921; Department of Biochemistry, Wolfson Laboratory, Imperial College of Science, Technology and Medicine, London SW7 2AY, England2; and Department of Pathology, Johns Hopkins University, Baltimore, Maryland 212053

Received 15 December 2000/Accepted 8 February 2001

BPHE-1 cells, which harbor 50 to 200 viral episomes, encapsidate viral genome and generate infectious bovine papillomavirus type 1 (BPV1) upon coexpression of capsid proteins L1 and L2 of BPV1, but not coexpression of BPV1 L1 and human papillomavirus type 16 (HPV16) L2. BPV1 L2 bound in vitro via its C-terminal 85 residues to purified L1 capsomers, but not with intact L1 virus-like particles in vitro. However, when the efficiency of BPV1 L1 coimmunoprecipitation with a series of BPV1 L2 deletion mutants was examined in vivo, the results suggested that residues 129 to 246 and 384 to 460 contain independent L1 interaction domains. An L2 mutant lacking the C-terminal L1 interaction domain was impaired for encapsidation of the viral genome. Coexpression of BPV1 L1 and a chimeric L2 protein composed of HPV16 L2 residues 1 to 98 fused to BPV1 L2 residues 99 to 469 generated infectious virions. However, inefficient encapsidation was seen when L1 was coexpressed with either BPV1 L2 with residues 91 to 246 deleted or with BPV1 L2 with residues 1 to 225 replaced with HPV16 L2. Impaired genome encapsidation did not correlate closely with impairment of the L2 proteins either to localize to promyelocytic leukemia oncogenic domains (PODs) or to induce localization of L1 or E2 to PODs. We conclude that the L1-binding domain located near the C terminus of L2 may bind L1 prior to completion of capsid assembly, and that both L1-binding domains of L2 are required for efficient encapsidation of the viral genome.

* Corresponding author. Mailing address: Department of Pathology, Room 656, The Ross Research Building, 720 Rutland Ave., Baltimore, MD 21205. Phone: (410) 502-5161. Fax: (410) 614-3548. E-mail: roden{at}jhmi.edu.

Journal of Virology, May 2001, p. 4332-4342, Vol. 75, No. 9
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.9.4332-4342.2001


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