Translation attenuation by PERK balances ER glycoprotein synthesis with lipid-linked oligosaccharide flux.
Endoplasmic reticulum (ER) homeostasis requires transfer and subsequent processing of the glycan Glc(3)Man(9)GlcNAc(2) (G(3)M(9)Gn(2)) from the lipid-linked oligosaccharide (LLO) glucose(3)mannose(9)N-acetylglucosamine(2)-P-P-dolichol (G(3)M(9)Gn(2)-P-P-Dol) to asparaginyl residues of nascent glycoprotein precursor polypeptides. However, it is unclear how the ER is protected against dysfunction from abnormal accumulation of LLO intermediates and aberrant N-glycosylation, as occurs in certain metabolic diseases. In metazoans phosphorylation of eukaryotic initiation factor 2alpha (eIF2alpha) on Ser(51) by PERK (PKR-like ER kinase), which is activated by ER stress, attenuates translation initiation. We use brief glucose deprivation to simulate LLO biosynthesis disorders, and show that attenuation of polypeptide synthesis by PERK promotes extension of LLO intermediates to G(3)M(9)Gn(2)-P-P-Dol under these substrate-limiting conditions, as well as counteract abnormal N-glycosylation. This simple mechanism requires eIF2alpha Ser(51) phosphorylation by PERK, and is mimicked by agents that stimulate cytoplasmic stress-responsive Ser(51) kinase activity. Thus, by sensing ER stress from defective glycosylation, PERK can restore ER homeostasis by balancing polypeptide synthesis with flux through the LLO pathway.
Shang
Jie
J
Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Gao
Ningguo
N
Kaufman
Randal J
RJ
Ron
David
D
Harding
Heather P
HP
Lehrman
Mark A
MA
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003513:Cycloheximide / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004789:Enzyme Activation; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis*; D006031:Glycosylation; D006706:Homeostasis; D008070:Lipopolysaccharides / Q000378:metabolism*; D010766:Phosphorylation; D014176:Protein Biosynthesis / Q000502:physiology*; D019892:eIF-2 Kinase / Q000502:physiology*
D003513:Cycloheximide; D015852:Eukaryotic Initiation Factor-2; D006023:Glycoproteins; D008070:Lipopolysaccharides; D019892:eIF-2 Kinase; C023023:lipid-linked oligosaccharides