Characterization of novel populations of MVM virions containing covalent DNA-protein complexes.
Virions of minute virus of mice were purified by sedimentation in sucrose gradients and chromatography on DEAE-cellulose columns and shown to consist of single-stranded viral DNA and the viral capsid polypeptides VP-1 (83 kDa) and VP-2 (64.5 kDa). A 63-kDa polypeptide distinct from the viral capsid polypeptide VP-3 (61.4 kDa) was found in some virion preparations. Virions sedimented at 135 and 110 S. The genomic single strands associated with purified 135 and 110 S virions were covalently bound to a protein as judged by the anomalous electrophoretic mobility of the DNA in agarose gels at pH 12.5. The protein was removed from the DNA by Pronase but remained bound after heating at 98 degrees in the presence of 0.1% sodium dodecyl sulfate. Nuclease digestion of the purified DNA-protein complex released several polypeptides ranging in size from 58 to 65 kDa. Restriction enzyme analysis of the purified DNA protein complex following its conversion to a duplex RF DNA in vitro showed that the protein was attached to the 5' termini of the DNA.
Faust
E A
EA
Cancer Research Laboratory, University of Western Ontario, London, Canada.
Brudzynska
K
K
Morgan
J
J
D000818:Animals; D002213:Capsid / Q000032:analysis; D002286:Carcinoma, Ehrlich Tumor; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D004277:DNA, Single-Stranded / Q000032:analysis*; D004279:DNA, Viral / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008915:Minute Virus of Mice / Q000032:analysis* / Q000235:genetics; D010321:Parvoviridae / Q000032:analysis*; D010455:Peptides / Q000032:analysis; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000032:analysis*; D014771:Virion / Q000032:analysis* / Q000235:genetics
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D010455:Peptides; D014764:Viral Proteins