Authors: Terenzi, Fulvia; deVeer, Michael J.; Ying, Han; Restifo, Nicholas P.; Williams, Bryan R. G.; Silverman, Robert H.

Source: Nucleic Acids Research, Volume 27, Number 22, OCTOBER 1999 , pp. 4369-4375(7)

Publisher: Oxford University Press

Abstract:

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.

Document Type: Research article

Affiliations: 1: Division of Clinical Sciences, Surgery Branch, Building 10, Room 2B42, National Cancer Institute, National Institutes of Health, 10 Center Drive, MSC 1502, Bethesda, MD 20892-1502, USA

Publication date: 1999-10-01

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• Nucleic Acids Research (NAR) is a fully Open Access journal, providing rapid publication of leading edge research into the nucleic acids under the following categories: chemistry, computational biology, genomics, molecular biology, nucleic acid enzymes, RNA and structural biology. There is a Survey and Summary section, and methods papers are published
  in NAR Methods Online. Each year the first issue is devoted to biological databases, and a later issue to relevant web-based software resources.

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