Inactivation of the Carney complex gene 1 (PRKAR1A) alters spatiotemporal regulation of cAMP and cAMP-dependent protein kinase: a study using genetically encoded FRET-based reporters

被引:17
作者
Cazabat, Laure [1 ,2 ,3 ]
Ragazzon, Bruno [1 ,2 ,3 ]
Varin, Audrey [4 ,5 ]
Potier-Cartereau, Marie [6 ,7 ]
Vandier, Christophe [6 ,7 ]
Vezzosi, Delphine [1 ,2 ,3 ]
Risk-Rabin, Marthe [1 ,2 ]
Guellich, Aziz [4 ,5 ]
Schittl, Julia [4 ,5 ]
Lechene, Patrick [4 ,5 ]
Richter, Wito [8 ]
Nikolaev, Viacheslav O. [9 ]
Zhang, Jin [10 ]
Bertherat, Jerome [1 ,2 ,3 ,11 ]
Vandecasteele, Gregoire [4 ,5 ]
机构
[1] INSERM, U1016, F-75014 Paris, France
[2] CNRS, UMR 8104, F-75014 Paris, France
[3] Univ Paris 05, F-75014 Paris, France
[4] INSERM, UMR S769, LabEx LERMIT, DHU TORINO, F-92296 Chatenay Malabry, France
[5] Univ Paris 11, Fac Pharm, F-92296 Chatenay Malabry, France
[6] INSERM, UMR1069, Tours, France
[7] Univ Tours, F-37032 Tours, France
[8] Univ Calif San Francisco, Dept Obstet Gynecol & Reprod Sci, San Francisco, CA 94143 USA
[9] Univ Gottingen, Med Ctr, D-37075 Gottingen, Germany
[10] Johns Hopkins Univ, Sch Med, Dept Pharmacol & Mol Sci, Baltimore, MD 21205 USA
[11] Hop Cochin, AP HP, Dept Endocrinol, Reference Ctr Rare Adrenal Dis, F-75674 Paris, France
基金
美国国家卫生研究院;
关键词
NODULAR ADRENOCORTICAL DISEASE; SUBUNIT TYPE 1A; ANCHORING PROTEIN; HORMONE RESISTANCE; CYCLIC-AMP; RI-ALPHA; PLASMA-MEMBRANE; EPAC ACTIVATION; ADRENAL-CORTEX; PKA-RI;
D O I
10.1093/hmg/ddt510
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
Carney complex (CNC) is a hereditary disease associating cardiac myxoma, spotty skin pigmentation and endocrine overactivity. CNC is caused by inactivating mutations in the PRKAR1A gene encoding PKA type I alpha regulatory subunit (RI alpha). Although PKA activity is enhanced in CNC, the mechanisms linking PKA dysregulation to endocrine tumorigenesis are poorly understood. In this study, we used Forster resonance energy transfer (FRET)-based sensors for cAMP and PKA activity to define the role of RI alpha in the spatiotemporal organization of the cAMP/PKA pathway. RI alpha knockdown in HEK293 cells increased basal as well as forskolin or prostaglandin E-1 (PGE(1))-stimulated total cellular PKA activity as reported by western blots of endogenous PKA targets and the FRET-based global PKA activity reporter, AKAR3. Using variants of AKAR3 targeted to subcellular compartments, we identified similar increases in the response to PGE(1) in the cytoplasm and at the outer mitochondrial membrane. In contrast, at the plasma membrane, the response to PGE(1) was decreased along with an increase in basal FRET ratio. These results were confirmed by western blot analysis of basal and PGE(1)-induced phosphorylation of membrane-associated vasodilator-stimulated phosphoprotein. Similar differences were observed between the cytoplasm and the plasma membrane in human adrenal cells carrying a RI alpha inactivating mutation. RI alpha inactivation also increased cAMP in the cytoplasm, at the outer mitochondrial membrane and at the plasma membrane, as reported by targeted versions of the cAMP indicator Epac1-camps. These results show that RI alpha inactivation leads to multiple, compartment-specific alterations of the cAMP/PKA pathway revealing new aspects of signaling dysregulation in tumorigenesis.
引用
收藏
页码:1163 / 1174
页数:12
相关论文
共 68 条
[1]   Subcellular dynamics of protein kinase A activity visualized by FRET-based reporters [J].
Allen, Michael D. ;
Zhang, Jin .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2006, 348 (02) :716-721
[2]   Protein kinase A regulates caspase-1 via Ets-1 in bone stromal cell-derived lesions: a link between cyclic AMP and pro-inflammatory pathways in osteoblast progenitors [J].
Almeida, Madson Q. ;
Tsang, Kit Man ;
Cheadle, Chris ;
Watkins, Tonya ;
Grivel, Jean-Charles ;
Nesterova, Maria ;
Goldbach-Mansky, Raphaela ;
Stratakis, Constantine A. .
HUMAN MOLECULAR GENETICS, 2011, 20 (01) :165-175
[3]   The essential role of RIα in the maintenance of regulated PKA activity [J].
Amieux, PS ;
McKnight, GS .
PROTEIN KINASE A AND HUMAN DISEASE, 2002, 968 :75-95
[4]   Increased basal cAMP-dependent protein kinase activity inhibits the formation of mesoderm-derived structures in the developing mouse embryo [J].
Amieux, PS ;
Howe, DG ;
Knickerbocker, H ;
Lee, DC ;
Su, T ;
Laszlo, GS ;
Idzerda, RL ;
McKnight, GS .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (30) :27294-27304
[5]   ACTIVATION OF PROTEIN KINASE BY PHYSIOLOGICAL CONCENTRATIONS OF CYCLIC-AMP [J].
BEAVO, JA ;
BECHTEL, PJ ;
KREBS, EG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1974, 71 (09) :3580-3583
[6]   Carney complex (CNC) [J].
Bertherat, Jerome .
ORPHANET JOURNAL OF RARE DISEASES, 2006, 1 (1)
[7]   PDE4D and PDE4B Function in Distinct Subcellular Compartments in Mouse Embryonic Fibroblasts [J].
Blackman, Brigitte E. ;
Horner, Kathleen ;
Heidmann, Julia ;
Wang, Dan ;
Richter, Wito ;
Rich, Thomas C. ;
Conti, Marco .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2011, 286 (14) :12590-12601
[8]   A guided tour into subcellular colocalization analysis in light microscopy [J].
Bolte, S. ;
Cordelieres, F. P. .
JOURNAL OF MICROSCOPY, 2006, 224 (213-232) :213-232
[9]   A Small Novel A-Kinase Anchoring Protein (AKAP) That Localizes Specifically Protein Kinase A-Regulatory Subunit I (PKA-RI) to the Plasma Membrane [J].
Burgers, Pepijn P. ;
Ma, Yuliang ;
Margarucci, Luigi ;
Mackey, Mason ;
van der Heyden, Marcel A. G. ;
Ellisman, Mark ;
Scholten, Arjen ;
Taylor, Susan S. ;
Heck, Albert J. R. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2012, 287 (52) :43789-43797
[10]   Haploinsufficiency at the protein kinase a RIα gene locus leads to fertility defects in male mice and men [J].
Burton, Kimberly A. ;
McDermott, Deborah A. ;
Wilkes, David ;
Poulsen, Melissa N. ;
Nolan, Michael A. ;
Goldstein, Marc ;
Basson, Craig T. ;
McKnight, G. Stanley .
MOLECULAR ENDOCRINOLOGY, 2006, 20 (10) :2504-2513