Activation of double-stranded RNA-activated protein kinase by mild impairment of oxidative metabolism in neurons.
Thiamine (vitamin B1) deficiency (TD) causes mild and chronic impairment of oxidative metabolism and induces neuronal death in specific brain regions. The mechanisms underlying TD-induced cell death, however, remain unclear. The double-stranded RNA-activated protein kinase (PKR), has been well known for its anti-viral function. Upon activation by viral infection or double-stranded RNA, PKR phosphorylates its substrate, the alpha-subunit of eukaryotic initiation factor-2 (eIF2alpha), leading to inhibition of translation. In response to various cellular stresses, PKR can also be stimulated by its protein activators, or its mouse homologue, PKR activator (RAX). We demonstrated that TD in mice induced phosphorylation of PKR at Thr446 and Thr451 and phosphorylation of eIF2alpha at Ser51 in the cerebellum and the thalamus. TD caused phosphorylation of PKR and eIF2alpha, as well as nuclear translocation of PKR in primary cultures of cerebellar granule neurons. PKR phosphorylation is necessary for its nuclear translocation because TD failed to induce nuclear translocation of a T446A/T451A PKR mutant. Both PKR inhibitor and dominant-negative PKR mutant protected cerebellar granule neurons against TD-induced cell death. TD promoted the association between RAX and PKR. Antioxidant vitamin E dramatically decreased the RAX/PKR association and ameliorated TD-induced cell death. Our results indicate that TD-induced neuronal death is at least partially mediated by the activation of PKR.
Wang
Xin
X
Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Fan
Zhiqin
Z
Wang
Bingwei
B
Luo
Jia
J
Ke
Zun-Ji
ZJ
D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D000975:Antioxidants / Q000378:metabolism / Q000494:pharmacology; D001921:Brain / Q000378:metabolism* / Q000503:physiopathology; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000502:physiology; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009410:Nerve Degeneration / Q000209:etiology / Q000378:metabolism* / Q000503:physiopathology; D009474:Neurons / Q000378:metabolism; D010085:Oxidative Phosphorylation; D018384:Oxidative Stress / Q000502:physiology*; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D012694:Serine / Q000378:metabolism; D013832:Thiamine Deficiency / Q000378:metabolism* / Q000503:physiopathology; D013912:Threonine / Q000378:metabolism; D024505:Tocopherols / Q000378:metabolism / Q000494:pharmacology; D019892:eIF-2 Kinase / Q000235:genetics / Q000378:metabolism*
D000975:Antioxidants; D015852:Eukaryotic Initiation Factor-2; D013912:Threonine; D012694:Serine; D019892:eIF-2 Kinase; D024505:Tocopherols