TY - JOUR
T1 - The antiviral enzymes PKR and RNase L suppress gene expression from viral and non-viral based vectors
AU - Terenzi, Fulvia
AU - DeVeer, Michael J.
AU - Ying, Han
AU - Restifo, Nicholas P.
AU - Williams, Bryan R.G.
AU - Silverman, Robert H.
PY - 1999/11/15
Y1 - 1999/11/15
N2 - Expression of transfected genes is shown to be suppressed by two intracellular enzymes, RNase L and protein kinase PKR, which function in interferon-treated cells to restrict viral replication. RNase L(-/-) or PKR(-/-) murine embryonic fibroblasts produced enhanced levels of protein from transfected genes compared with wild-type cells. Increased expression of exogenous genes in RNase L(-/-) cells correlated with elevated levels of mRNA and thus appeared to be due to enhanced mRNA stability. Plasmid encoding adenovirus VA RNAs was able to further enhance accumulation of the exogenous gene transcript and protein, even in cells lacking PKR. In contrast to the increased expression of transfected genes in cells lacking RNase L or PKR, expression of endogenous host genes was unaffected by the absence of these enzymes. In addition, a dominant-negative PKR mutant improved expression from a conventional plasmid vector and from a Semliki Forest virus derived, self-replicating vector. These results indicate that viral infections and transfections produce similar stress responses in mammalian cells and suggest strategies for selectively increasing expression of exogenous genes.
AB - Expression of transfected genes is shown to be suppressed by two intracellular enzymes, RNase L and protein kinase PKR, which function in interferon-treated cells to restrict viral replication. RNase L(-/-) or PKR(-/-) murine embryonic fibroblasts produced enhanced levels of protein from transfected genes compared with wild-type cells. Increased expression of exogenous genes in RNase L(-/-) cells correlated with elevated levels of mRNA and thus appeared to be due to enhanced mRNA stability. Plasmid encoding adenovirus VA RNAs was able to further enhance accumulation of the exogenous gene transcript and protein, even in cells lacking PKR. In contrast to the increased expression of transfected genes in cells lacking RNase L or PKR, expression of endogenous host genes was unaffected by the absence of these enzymes. In addition, a dominant-negative PKR mutant improved expression from a conventional plasmid vector and from a Semliki Forest virus derived, self-replicating vector. These results indicate that viral infections and transfections produce similar stress responses in mammalian cells and suggest strategies for selectively increasing expression of exogenous genes.
UR - http://www.scopus.com/inward/record.url?scp=0033571204&partnerID=8YFLogxK
U2 - 10.1093/nar/27.22.4369
DO - 10.1093/nar/27.22.4369
M3 - Article
C2 - 10536144
AN - SCOPUS:0033571204
SN - 0305-1048
VL - 27
SP - 4369
EP - 4375
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 22
ER -