DNA encapsidation by viruslike particles assembled in insect cells from the major capsid protein VP1 of B-lymphotropic papovavirus.
Capsids of polyomaviruses--small, nonenveloped DNA viruses--consist of the major structural protein VP1 and the minor structural proteins VP2 and VP3. The contributions of the individual capsid proteins to functions of the viral particle, such as DNA encapsidation, cell receptor attachment, entry, and uncoating, are still not clear. Here we show that viruslike particles assembled in nuclei of insect cells from VP1 of the monkey B-lymphotropic papovavirus (LPV) are sufficient to unspecifically encapsidate DNA. LPV VP1 expressed in large amounts in insect cells by a baculovirus vector assembled spontaneously in the nuclei to form viruslike particles. After metrizamide equilibrium density gradient purification and nuclease digestion, a fraction of these particles was shown to contain VP1-associated linear, double-stranded DNA with a predominant size of 4.5 kb. The fraction of DNA-containing VP1 particles increased with time and dose of baculovirus vector infection. The DNA-containing particles, further purified by sucrose gradient centrifugation, appeared as "full" particles in negative-staining electron microscopy. As shown by DNA hybridization, the encapsidated DNA consisted of insect cell and baculoviral sequences with no apparent strong homology to LPV sequences. Three non-LPV VP1-derived host proteins with apparent molecular masses of approximately 14, 15, and 16 kDa copurified with the DNA-containing particles and may represent insect cell histones encapsidated together with the DNA. A similar species of host DNA was also found in purified LPV wild-type virions. These data suggest that LPV VP1 alone can be sufficient to encapsidate linear DNA in a sequence-independent manner.
Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, Heidelberg, Germany. M.Pawlita@dkfz-heidelberg.de.
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Pawlita
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Müller
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Oppenländer
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Zentgraf
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Herrmann
D000818:Animals; D001402:B-Lymphocytes / Q000821:virology; D002213:Capsid / Q000235:genetics / Q000502:physiology*; D036022:Capsid Proteins; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism*; D006657:Histones; D016393:Lymphoma, B-Cell / Q000473:pathology; D027383:Papillomaviridae / Q000502:physiology*; D027382:Polyomaviridae; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D018411:Spodoptera / Q000166:cytology; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014771:Virion; D019065:Virus Assembly
D036022:Capsid Proteins; D004279:DNA, Viral; D006657:Histones; D011993:Recombinant Fusion Proteins; D004247:DNA