Cell Signaling Technology

Product Pathways - Translational Control

GCN2 Antibody #3302

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk Endogenous 220 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

GCN2 Antibody detects endogenous levels of GCN2 protein independent of phosphorylation.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to sequence near the amino terminus of human GCN2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from ME180 and HT1376 cells that were untreated, treated with UV light (50mJ/cm2, 30 minutes), or subjected to nocodazole block (50 ng/ml, 24hrs), using GCN2 Antibody.

Background

Phosphorylation of the alpha subunit of eukaryotic initiation factor 2 is a well documented mechanism of downregulating protein synthesis under a variety of stress conditions. Kinases activated by viral infection (PKR), endoplasmic reticulum stress (PERK/PEK), amino acid deprivation (GCN2) and hemin deficiency (HRI) can phosphorylate the alpha subunit of eIF2 (1,2). GCN2 is also required for UV-light induced translation inhibition, and in vivo phosphorylation of murine GCN2 at Thr898 is induced by both UV irradiation and by leucine deprivation (3). UV-induced activation of NF-kappaB also requires GCN2, which may act simply by preventing translation of IkappaB-alpha to replace pools that have been ubiquitinated and degraded (4). Interestingly, proteasome inhibitors (MG132 and ALLN) activate the GCN2/eIF2alpha pathway, suggesting a pivotal role for this kinase in stress response and ubiquitin-mediated signaling (5). In vitro autophosphorylation of yeast GCN2 within its activation loop at Thr882 and Thr887 (Thr898 and Thr903 in mouse) has also been reported (6).

  1. Kaufman, R.J. (1999) Genes Dev. 13, 1211-1233.
  2. Sheikh, M.S. and Fornace, A.J. (1999) Oncogene 18, 6121-6128.
  3. Deng, J. et al. (2002) Curr. Biol. 12, 1279-1286.
  4. Jiang, H.Y. and Wek, R.C. (2005) Biochem. J. 385, 371-380.
  5. Jiang, H.Y. and Wek, R.C. (2005) J. Biol. Chem. XXX, XXX-XXX.
  6. Garcia-Barrio, M. et al. (2002) J. Biol. Chem. 277, 30675-30683.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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