Relative transcriptional inducibility of the human interferon-alpha subtypes conferred by the virus-responsive enhancer sequence.
This paper addresses the role of transcriptional regulation in the determination of the levels of expression of different interferon-alpha subtypes secreted from Namalwa cells following infection with Sendai virus. Using RT-PCR to determine the relative abundance of mRNA species coding for the various subtypes, we found a general correlation with corresponding protein levels, indicative of a role for transcriptional control in the determination of levels of individual subtypes. We have used reporter gene constructs to compare the inducibility of the virus-response elements from the IFNA1, A2, A4, and A14 subtype genes cloned upstream of a secreted alkaline phosphatase gene. The inducibility of these reporter gene constructs broadly correlated with the relative mRNA abundances in both transiently and stably transfected Namalwa cells. During work with stable cell lines, we found that G418, the drug used for the selection of transfected cells, inhibited the induction of interferon by both Sendai virus and double-stranded RNA. This inhibition was reversible when G418 was removed from the medium 24 h before the addition of virus.
Dent
C L
CL
Division of Biology, Wellcome Research Labs, Beckenham, Kent, UK.
Macbride
S J
SJ
Sharp
N A
NA
Gewert
D R
DR
D001483:Base Sequence; D002460:Cell Line; D004742:Enhancer Elements, Genetic; D017930:Genes, Reporter; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics*; D008969:Molecular Sequence Data; D010222:Parainfluenza Virus 1, Human; D018184:Paramyxoviridae Infections / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods; D012194:RNA-Directed DNA Polymerase; D015203:Reproducibility of Results; D014158:Transcription, Genetic
D016898:Interferon-alpha; D012194:RNA-Directed DNA Polymerase