Expression of vaccinia virus K3L protein in yeast inhibits eukaryotic initiation factor-2 kinase GCN2 and the general amino acid control pathway

Weijun Qian, Shuhao Zhu, Alexander Y. Sobolev, Ronald Wek

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Abstract

Phosphorylation of the α subunit of eukaryotic initiation factor-2 (eIF- 2) is a well characterized mechanism regulating protein synthesis. Viral and cellular proteins have been identified that regulate the activity of the eIF- 2α kinases. The regulatory protein, K3L, from vaccinia virus is homologous to the amino terminus of eIF-2α and is thought to inhibit the activity of the double-stranded RNA-dependent kinase suppressing the antiviral mechanism mediated by this kinase. We investigated whether K3L can inhibit the activity of the yeast eIF-2α kinase GCN2. Expression of K3L protein in yeast reduced the level of eIF-2α phosphorylation by GCN2 and blocked the stimulation of the general amino acid control pathway in response to starvation conditions. Accompanying in vitro studies showed that recombinant K3L protein reduced GCN2 autophosphorylation and phosphorylation eIF-2α. In agreement with the hypothesis that K3L inhibits eIF-2α kinases by functioning as a pseudosubstrate, we observed that K3L directly interacted with the kinase catalytic domain of GCN2. Together, these results indicate that K3L is a specific inhibitor of eIF-2α kinases from mammals and yeast and suggest that the kinases contain common structural features important for recognition of their substrate eIF-2α.

Original languageEnglish
Pages (from-to)13202-13207
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number22
DOIs
StatePublished - 1996

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Eukaryotic Initiation Factor-2
Yeast
eIF-2 Kinase
Phosphotransferases
Yeasts
Phosphorylation
Amino Acids
Recombinant proteins
Proteins
Mammals
Fungal Proteins
Double-Stranded RNA
Viral Proteins
Starvation
Recombinant Proteins
Antiviral Agents
Catalytic Domain
Vaccinia virus pK3 protein
Substrates

ASJC Scopus subject areas

  • Biochemistry

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Expression of vaccinia virus K3L protein in yeast inhibits eukaryotic initiation factor-2 kinase GCN2 and the general amino acid control pathway. / Qian, Weijun; Zhu, Shuhao; Sobolev, Alexander Y.; Wek, Ronald.

In: Journal of Biological Chemistry, Vol. 271, No. 22, 1996, p. 13202-13207.

Research output: Contribution to journalArticle

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AU - Wek, Ronald

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