Defect in entry and altered pathogenicity of a polyoma virus mutant blocked in VP2 myristylation.
We have examined effects of mutations of the minor capsid proteins VP2 and VP3 of polyoma virus. Infection of cells by mutants blocked in expression of either VP2 or VP3 by alterations of their initiator methionine codons led to the rapid selection of wild-type revertants. This reversion suggests a strong, if not absolute, requirement for both proteins in virus growth. A mutant virus, VP2*, in which alanine was substituted for glycine-2, expressed a nonmyristylated form of VP2 that was incorporated into virions. Studies of VP2* revealed a 15- to 20-fold lower specific infectivity and a delay in growth in both primary and established mouse cells compared to wild-type virus. Analysis of the growth delay indicated a defect in an early step of infection, at or prior to uncoating. Synthesis of viral DNA and VP1 was delayed by about 9 hr in mutant compared to that in wild-type infected cells. No evidence was obtained for an effect on either virus assembly (encapsidation) or stability of virions. Infection of mice by the VP2* mutant virus resulted in attenuated virus replication and tumor induction which were much more severely affected following intranasal inoculation than intraperitoneal inoculation.
Department of Pathology, Harvard Medical School, Boston, MA 02115.
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R
Sahli
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R
Freund
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Dubensky
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Garcea
R
R
Bronson
T
T
Benjamin
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D002478:Cells, Cultured; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis; D006384:Hemagglutination; D006358:Hot Temperature; D066298:In Vitro Techniques; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009226:Myristates / Q000378:metabolism; D009374:Neoplasms, Experimental / Q000382:microbiology; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011120:Polyomavirus / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D011499:Protein Processing, Post-Translational; D051381:Rats; D014162:Transfection; D014779:Virus Replication
D036022:Capsid Proteins; D004279:DNA, Viral; D009226:Myristates; D009838:Oligodeoxyribonucleotides; C442543:VP2 protein, Polyomavirus