The synthesis of influenza virus negative-strand RNA takes place in insoluble complexes present in the nuclear matrix fraction.
The replication of influenza virus RNA in vitro has been studied by cell fractionation of MDCK-infected cells and characterization of in vitro synthesized RNA. Analysis of the RNA product polarity by liquid hybridization to excess single-stranded DNA probes shows that only the RNP complexes present in the nuclear matrix fraction are able to synthesize negative-polarity RNA. This RNA product has been characterized as authentic vRNA by size analysis, RNase-protection by unlabelled, positive-polarity riboprobes and T1-fingerprinting. Priming the in vitro reaction with ApG stimulates preferentially the synthesis of positive-polarity RNA, while ApGpU stimulates both positive and negative-polarity RNA synthesis.
López-Turiso
J A
JA
Centro de Biologia Molecular (CSIC-UAM), Universidad Autónoma, Canto Blanco, Madrid, Spain.
Martínez
C
C
Tanaka
T
T
Ortín
J
J
D000818:Animals; D002458:Cell Fractionation; D002460:Cell Line; D003001:Cloning, Molecular; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D009980:Influenza A virus / Q000201:enzymology / Q000235:genetics*; D015530:Nuclear Matrix / Q000382:microbiology*; D009693:Nucleic Acid Hybridization; D009843:Oligoribonucleotides; D012367:RNA, Viral / Q000096:biosynthesis* / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism; D012261:Ribonucleoproteins
D015342:DNA Probes; D009843:Oligoribonucleotides; D012367:RNA, Viral; D012261:Ribonucleoproteins; D012324:RNA-Dependent RNA Polymerase