Translational regulation by the interferon-induced double-stranded-RNA-activated 68-kDa protein kinase

Glen N Barber, Marlene Wambach, Min Liang Wong, Thomas E. Dever, Alan G. Hinnebusch, Michael G. Katze

Research output: Contribution to journalArticle

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Abstract

Activation of the interferon-inducible 68-kDa protein kinase (referred to as P68) by double-stranded RNA catalyzes phosphorylation of the α subunit of eukaryotic protein synthesis initiation factor 2. We have analyzed the transient expression of mutant and wild-type kinase molecules in transfected COS cells to examine the effects of the kinase on gene expression in the absence of other interferon-induced gene products. The wild-type P68 kinase was expressed inefficiently whereas a catalytically inactive P68 was expressed at 30- to 40-fold higher levels. Protein stability measurements and primer-extension analysis of human kinase-specific mRNA levels provided evidence that kinase expression was regulated at the level of mRNA translation. Further, cotransfection experiments revealed that the domain II catalytically inactive mutant could stimulate reporter gene protein synthesis in a transdominant manner. We also examined the expression of mutants with deletions in the N-terminal double-stranded RNA binding domains and found that a kinase construct lacking aa 156-243 was expressed at levels comparable to the wild type whereas a P68 construct lacking aa 91-243 was expressed at levels 70-fold higher. Both the inactive domain II P68 mutant and the deletion mutant lacking aa 91-243 were less inhibitory to growth in yeast due to the reduced ability to phosphorylate initiation factor 2α in vivo. In conclusion we have demonstrated that the P68 kinase can regulate mRNA translation primarily of its own mRNA and to a lesser extent of a heterologous mRNA and that this regulation is notably affected by mutations in either the catalytic or N-terminal regulatory domains.

Original languageEnglish
Pages (from-to)4621-4625
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number10
StatePublished - May 15 1993
Externally publishedYes

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Double-Stranded RNA
Protein Kinases
Interferons
Phosphotransferases
Prokaryotic Initiation Factor-2
eIF-2 Kinase
Protein Biosynthesis
Messenger RNA
Protein Stability
COS Cells
Protein Subunits
Reporter Genes
Yeasts
Phosphorylation
Gene Expression
Mutation
Growth
Genes
Proteins

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Translational regulation by the interferon-induced double-stranded-RNA-activated 68-kDa protein kinase. / Barber, Glen N; Wambach, Marlene; Wong, Min Liang; Dever, Thomas E.; Hinnebusch, Alan G.; Katze, Michael G.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 90, No. 10, 15.05.1993, p. 4621-4625.

Research output: Contribution to journalArticle

Barber, Glen N ; Wambach, Marlene ; Wong, Min Liang ; Dever, Thomas E. ; Hinnebusch, Alan G. ; Katze, Michael G. / Translational regulation by the interferon-induced double-stranded-RNA-activated 68-kDa protein kinase. In: Proceedings of the National Academy of Sciences of the United States of America. 1993 ; Vol. 90, No. 10. pp. 4621-4625.
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