Activation of Dual Oxidases Duox1 and Duox2 DIFFERENTIAL REGULATION MEDIATED BY cAMP-DEPENDENT PROTEIN KINASE AND PROTEIN KINASE C-DEPENDENT PHOSPHORYLATION

被引:163
作者
Rigutto, Sabrina [1 ]
Hoste, Candice [1 ]
Grasberger, Helmut [3 ]
Milenkovic, Milutin [1 ]
Communi, David [1 ]
Dumont, Jacques E. [1 ]
Corvilain, Bernard [1 ,2 ]
Miot, Francoise [1 ]
De Deken, Xavier [1 ]
机构
[1] Univ Libre Bruxelles, IRIBHM, Hop Erasme, B-1070 Brussels, Belgium
[2] Univ Libre Bruxelles, Dept Endocrinol, Hop Erasme, B-1070 Brussels, Belgium
[3] Univ Chicago, Dept Med, Chicago, IL 60637 USA
关键词
HUMAN THYROID-CELLS; CONGENITAL HYPOTHYROIDISM; NADPH OXIDASE; HYDROGEN-PEROXIDE; H2O2-GENERATING SYSTEM; MATURATION FACTOR; H2O2; GENERATION; TSH RECEPTOR; GENE; MUTATIONS;
D O I
10.1074/jbc.M806893200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Dual oxidases were initially identified as NADPH oxidases producing H2O2 necessary for thyroid hormone biosynthesis. The crucial role of Duox2 has been demonstrated in patients suffering from partial iodide organification defect caused by bi-allelic mutations in the DUOX2 gene. However, the Duox1 function in thyroid remains elusive. We optimized a functional assay by co-expressing Duox1 or Duox2 with their respective maturation factors, DuoxA1 and DuoxA2, to compare their intrinsic enzymatic activities under stimulation of the major signaling pathways active in the thyroid in relation to their membrane expression. We showed that basal activity of both Duox isoenzymes depends on calcium and functional EF-hand motifs. However, the two oxidases are differentially regulated by activation of intracellular signaling cascades. Duox1 but not Duox2 activity is stimulated by forskolin (EC50 = 0.1 mu M) via protein kinase A-mediated Duox1 phosphorylation on serine 955. In contrast, phorbol esters induce Duox2 phosphorylation via protein kinase C activation associated with high H2O2 generation (phorbol 12-myristate 13-acetate EC50 = 0.8 nM). These results were confirmed in human thyroid cells, suggesting that Duox1 is also involved in thyroid hormonogenesis. Our data provide, for the first time, detailed insights into the mechanisms controlling the activation of Duox1-2 proteins and reveal additional phosphorylation-mediated regulation.
引用
收藏
页码:6725 / 6734
页数:10
相关论文
共 51 条
[1]  
ALLGEIER A, 1994, J BIOL CHEM, V269, P13733
[2]   Dual oxidase-2 has an intrinsic Ca2+-dependent H2O2-generating activity [J].
Ameziane-El-Hassani, R ;
Morand, S ;
Boucher, JL ;
Frapart, YM ;
Apostolou, D ;
Agnandji, D ;
Gnidehou, S ;
Ohayon, R ;
Noël-Hudson, MS ;
Francon, J ;
Lalaoui, K ;
Virion, A ;
Dupuy, C .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (34) :30046-30054
[3]   NADPH oxidase: An update [J].
Babior, BM .
BLOOD, 1999, 93 (05) :1464-1476
[4]   NOX3, a superoxide-generating NADPH oxidase of the inner ear [J].
Bánfi, B ;
Malgrange, B ;
Knisz, J ;
Steger, K ;
Dubois-Dauphin, M ;
Krause, KH .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (44) :46065-46072
[5]   Mechanism of Ca2+ activation of the NADPH oxidase 5 (NOX5) [J].
Bánfi, B ;
Tirone, F ;
Durussel, I ;
Knisz, J ;
Moskwa, P ;
Molnár, GZ ;
Krause, KH ;
Cox, JA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (18) :18583-18591
[6]   Two novel proteins activate superoxide generation by the NADPH oxidase NOX1 [J].
Bánfi, B ;
Clark, RA ;
Steger, K ;
Krause, KH .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (06) :3510-3513
[7]  
BENARD B, 1971, UNION MED CAN, V100, P701
[8]   REGULATION OF GENE-EXPRESSION BY TRANSFECTED SUBUNITS OF CAMP-DEPENDENT PROTEIN-KINASE [J].
BUCHLER, W ;
MEINECKE, M ;
CHAKRABORTY, T ;
JAHNSEN, T ;
WALTER, U ;
LOHMANN, SM .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 188 (02) :253-259
[9]   Nox3 regulation by NOXO1, p47phox, and p67phox [J].
Cheng, GJ ;
Ritsick, D ;
Lambeth, JD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (33) :34250-34255
[10]   ROLE OF THE CYCLIC ADENOSINE-3',5'-MONOPHOSPHATE AND THE PHOSPHATIDYLINOSITOL CA2+ CASCADES IN MEDIATING THE EFFECTS OF THYROTROPIN AND IODIDE ON HORMONE SYNTHESIS AND SECRETION IN HUMAN THYROID SLICES [J].
CORVILAIN, B ;
LAURENT, E ;
LECOMTE, M ;
VANSANDE, J ;
DUMONT, JE .
JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM, 1994, 79 (01) :152-159