Structure and function of the polypeptides in simian virus 40. II. Transcription of subviral deoxynucleoprotein complexes in vitro.
A deoxynucleoprotein complex (DNP-1) isolated from simian virus 40 (SV40) after disruption of the virus in an alkaline buffer contains the viral deoxyribonucleic acid (DNA) and four minor structural polypeptides. Dissociation of DNP-I by equilibrium centrifugation in CsCl yields a complex (DNP-II) that contains a small amount of polypeptide tightly bound to the viral DNA. Studies of the template activity of these deoxynucleoprotein complexes in vitro with Escherichia coli transcriptase show that the rate of transcription of DNP-I and DNP-II is 30 and 80%, respectively, compared with that of deproteinized SV40 DNA component I. In dimethyl sulfoxide gradients, the complementary ribonucleic acid (cRNA) synthesized from DNP-I is one-third to one-half the size of the cRNA species from DNA-I and DNP-II. Competition hybridization experiments show that with the E. coli transcriptase only a portion (about one-half) of the SV40 genome is transcribed with DNP-I as template, whereas most or all of the genome is transcribed with DNP-II as template. The template activity of the deoxynucleoprotein complexes with a highly active form II ribonucleic acid polymerase prepared from SV40-infected permissive cells follows similar transcription kinetics. The results indicate that structural nucleoproteins of SV40 bind nonrandomly to the viral DNA and effect the transcription of some subset of its sequences in vitro.
D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004279:DNA, Viral; D004121:Dimethyl Sulfoxide; D004926:Escherichia coli / Q000201:enzymology; D005815:Genetic Code; D005827:Genetics, Microbial; D000882:Haplorhini; D007668:Kidney; D009693:Nucleic Acid Hybridization; D009698:Nucleoproteins / Q000096:biosynthesis*; D010452:Peptide Biosynthesis; D012316:RNA Nucleotidyltransferases / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis; D013539:Simian virus 40 / Q000378:metabolism*; D013698:Templates, Genetic; D014316:Tritium; D014500:Uracil Nucleotides / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis*
D004279:DNA, Viral; D009698:Nucleoproteins; D012367:RNA, Viral; D014500:Uracil Nucleotides; D014764:Viral Proteins; D014316:Tritium; D012316:RNA Nucleotidyltransferases; D004121:Dimethyl Sulfoxide