Identification of genes in thyrocytes regulated by unfolded protein response by using disulfide bond reducing agent of dithiothreitol.
Disulfide bonds are formed between the sulfhydryl groups in two cysteine residues of a protein. The formation of these bonds is necessary for the proper folding of a protein into its active three-dimensional form. In this study, the genes associated with disulfide bond formation of proteins from the rat thyroid cell line, FRTL-5 cell, were investigated using disulfide bond reducing agent of dithiothreitol (DTT), which prevented disulfide formation of newly synthesized proteins. The expression of six genes, they being the cAMP phosphodiesterase 7A1, neuronal cell death inducible putative kinase (NIPK), cytosolic LIM protein (Ajuba), Eker, early growth response 1 and the ferritin heavy chain, was specifically enhanced under both reductive conditions and various endoplasmic reticulum (ER) stresses inducing drugs such as Brefeldin A (BFA), calcium ionophore A23187 (A23187) and tunicamycin. These results suggest that a suitable redox environment is necessary for the correct disulfide bond conformation in thyrocytes in a complex system.
Department of Biology, Chungnam National University, Taejon, Korea.
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Park
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Hwang
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Shong
O Y
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Kwon
D000818:Animals; D015152:Blotting, Northern; D020126:Brefeldin A / Q000494:pharmacology; D000001:Calcimycin; D002460:Cell Line; D004220:Disulfides / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D018398:Homeodomain Proteins; D007476:Ionophores / Q000494:pharmacology; D060588:LIM Domain Proteins; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D017510:Protein Folding; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000235:genetics; D051381:Rats; D013961:Thyroid Gland / Q000378:metabolism / Q000502:physiology*; D014415:Tunicamycin / Q000494:pharmacology
C119385:Ajuba protein, rat; D004220:Disulfides; D018398:Homeodomain Proteins; D007476:Ionophores; D060588:LIM Domain Proteins; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D014415:Tunicamycin; D020126:Brefeldin A; D000001:Calcimycin; D012313:RNA; D004229:Dithiothreitol