Drosophila G-protein-coupled receptor kinase 2 regulates cAMP-dependent Hedgehog signaling

被引:12
作者
Cheng, Shuofei [1 ,2 ]
Maier, Dominic [1 ,2 ]
Hipfner, David R. [1 ,2 ,3 ]
机构
[1] Inst Rech Clin Montreal, Montreal, PQ H2W 1R7, Canada
[2] McGill Univ, Dept Anat & Cell Biol, Montreal, PQ H3A 2B2, Canada
[3] Univ Montreal, Dept Med, Montreal, PQ H3C 3J7, Canada
来源
DEVELOPMENT | 2012年 / 139卷 / 01期
基金
加拿大健康研究院;
关键词
Hedgehog; Smoothened; G-protein-coupled receptor kinase; cAMP; Drosophila; TARGET GENE-EXPRESSION; CUBITUS-INTERRUPTUS; SMOOTHENED PHOSPHORYLATION; REQUIRES PHOSPHORYLATION; LIMB DEVELOPMENT; TRANSDUCTION; PATHWAY; ROLES; MICE; DESENSITIZATION;
D O I
10.1242/dev.068817
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
G-protein-coupled receptor kinases (GRKs) play a conserved role in Hedgehog (Hh) signaling. In several systems, GRKs are required for efficient Hh target gene expression. Their principal target appears to be Smoothened (Smo), the intracellular signal-generating component of the pathway and a member of the G-protein-coupled receptor (GPCR) protein family. In Drosophila, a GRK called Gprk2 is needed for internalization and downregulation of activated Smo, consistent with the typical role of these kinases in negatively regulating GPCRs. However, Hh target gene activation is strongly impaired in gprk2 mutant flies, indicating that Gprk2 must also positively regulate Hh signaling at some level. To investigate its function in signaling, we analyzed several different readouts of Hh pathway activity in animals or cells lacking Gprk2. Surprisingly, although target gene expression was impaired, Smo-dependent activation of downstream components of the signaling pathway was increased in the absence of Gprk2. This suggests that Gprk2 does indeed play a role in terminating Smo signaling. However, loss of Gprk2 resulted in a decrease in cellular cAMP concentrations to a level that was limiting for Hh target gene activation. Normal expression of target genes was restored in gprk2 mutants by stimulating cAMP production or activating the cAMP-dependent Protein kinase A (Pka). Our results suggest that direct regulation of Smo by Gprk2 is not absolutely required for Hh target gene expression. Gprk2 is important for normal cAMP regulation, and thus has an indirect effect on the activity of Pka-regulated components of the Hh pathway, including Smo itself.
引用
收藏
页码:85 / 94
页数:10
相关论文
共 55 条
[21]   Use of a cAMP BRET sensor to characterize a novel regulation of cAMP by the sphingosine 1-phosphate/G13 pathway [J].
Jiang, Lily I. ;
Collins, Julie ;
Davis, Richard ;
Lin, Keng-Mean ;
DeCamp, Dianne ;
Roach, Tamara ;
Hsueh, Robert ;
Rebres, Robert A. ;
Ross, Elliott M. ;
Taussig, Ronald ;
Fraser, Iain ;
Sternweis, Paul C. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2007, 282 (14) :10576-10584
[22]   Increased acute inflammation, leukotriene B4-induced chemotaxis, and signaling in mice deficient for G protein-coupled receptor kinase 6 [J].
Kavelaars, A ;
Vroon, A ;
Raatgever, RP ;
Fong, AM ;
Premont, RT ;
Patel, DD ;
Lefkowitz, RJ ;
Heijnen, CJ .
JOURNAL OF IMMUNOLOGY, 2003, 171 (11) :6128-6134
[23]   Patched regulates Smoothened trafficking using lipoprotein-derived lipids [J].
Khaliullina, Helena ;
Panakova, Daniela ;
Eugster, Christina ;
Riedel, Falko ;
Carvalho, Maria ;
Eaton, Suzanne .
DEVELOPMENT, 2009, 136 (24) :4111-4121
[24]  
Klein RS, 2001, DEVELOPMENT, V128, P1971
[25]   Gprk2 controls cAMP levels in Drosophila development [J].
Lannutti, BJ ;
Schneider, LE .
DEVELOPMENTAL BIOLOGY, 2001, 233 (01) :174-185
[26]   Disruption of the PACAP gene promotes medulloblastoma in ptc1 mutant mice [J].
Lelievre, Vincent ;
Seksenyan, Akop ;
Nobuta, Hiroko ;
Yong, William H. ;
Chhith, Seririthanar ;
Niewiadomski, Pawel ;
Cohen, Joseph R. ;
Dong, Hongmei ;
Flores, Avegail ;
Liau, Linda M. ;
Kornblum, Harley I. ;
Scott, Matthew P. ;
Waschek, James A. .
DEVELOPMENTAL BIOLOGY, 2008, 313 (01) :359-370
[27]   FUNCTION OF PROTEIN KINASE-A IN HEDGEHOG SIGNAL-TRANSDUCTION AND DROSOPHILA IMAGINAL DISC DEVELOPMENT [J].
LI, W ;
OHLMEYER, JT ;
LANE, ME ;
KALDERON, D .
CELL, 1995, 80 (04) :553-562
[28]   Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells [J].
Lum, L ;
Yao, SQ ;
Mozer, B ;
Rovescalli, A ;
Von Kessler, D ;
Nirenberg, M ;
Beachy, PA .
SCIENCE, 2003, 299 (5615) :2039-2045
[29]   Regulation of mammalian Gli proteins by Costal 2 and PKA in Drosophila reveals Hedgehog pathway conservation [J].
Marks, Steven A. ;
Kalderon, Daniel .
DEVELOPMENT, 2011, 138 (12) :2533-2542
[30]   Smoothened signal transduction is promoted by G protein-coupled receptor kinase 2 [J].
Meloni, Alison R. ;
Fralish, Gregory B. ;
Kelly, Patrick ;
Salahpour, Ali ;
Chen, James K. ;
Wechsler-Reya, Robert J. ;
Lefkowitz, Robert J. ;
Caron, Marc G. .
MOLECULAR AND CELLULAR BIOLOGY, 2006, 26 (20) :7550-7560