Positional Cloning of a Type 2 Diabetes Quantitative Trait Locus; , a Negative Regulator of Insulin Secretion
We previously mapped a type 2 diabetes (T2D) locus on chromosome 16 (Chr 16) in an F2 intercross from the BTBR T (+) tf (BTBR) Lepob/ob and C57BL/6 (B6) Lepob/ob mouse strains. Introgression of BTBR Chr 16 into B6 mice resulted in a consomic mouse with reduced fasting plasma insulin and elevated glucose levels. We derived a panel of sub-congenic mice and narrowed the diabetes susceptibility locus to a 1.6 Mb region. Introgression of this 1.6 Mb fragment of the BTBR Chr 16 into lean B6 mice (B6.16BT36–38) replicated the phenotypes of the consomic mice. Pancreatic islets from the B6.16BT36–38 mice were defective in the second phase of the insulin secretion, suggesting that the 1.6 Mb region encodes a regulator of insulin secretion. Within this region, syntaxin-binding protein 5-like (Stxbp5l) or tomosyn-2 was the only gene with an expression difference and a non-synonymous coding single nucleotide polymorphism (SNP) between the B6 and BTBR alleles. Overexpression of the b-tomosyn-2 isoform in the pancreatic β-cell line, INS1 (832/13), resulted in an inhibition of insulin secretion in response to 3 mM 8-bromo cAMP at 7 mM glucose. In vitro binding experiments showed that tomosyn-2 binds recombinant syntaxin-1A and syntaxin-4, key proteins that are involved in insulin secretion via formation of the SNARE complex. The B6 form of tomosyn-2 is more susceptible to proteasomal degradation than the BTBR form, establishing a functional role for the coding SNP in tomosyn-2. We conclude that tomosyn-2 is the major gene responsible for the T2D Chr 16 quantitative trait locus (QTL) we mapped in our mouse cross. Our findings suggest that tomosyn-2 is a key negative regulator of insulin secretion.
Vyšlo v časopise:
Positional Cloning of a Type 2 Diabetes Quantitative Trait Locus; , a Negative Regulator of Insulin Secretion. PLoS Genet 7(10): e32767. doi:10.1371/journal.pgen.1002323
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002323
Souhrn
We previously mapped a type 2 diabetes (T2D) locus on chromosome 16 (Chr 16) in an F2 intercross from the BTBR T (+) tf (BTBR) Lepob/ob and C57BL/6 (B6) Lepob/ob mouse strains. Introgression of BTBR Chr 16 into B6 mice resulted in a consomic mouse with reduced fasting plasma insulin and elevated glucose levels. We derived a panel of sub-congenic mice and narrowed the diabetes susceptibility locus to a 1.6 Mb region. Introgression of this 1.6 Mb fragment of the BTBR Chr 16 into lean B6 mice (B6.16BT36–38) replicated the phenotypes of the consomic mice. Pancreatic islets from the B6.16BT36–38 mice were defective in the second phase of the insulin secretion, suggesting that the 1.6 Mb region encodes a regulator of insulin secretion. Within this region, syntaxin-binding protein 5-like (Stxbp5l) or tomosyn-2 was the only gene with an expression difference and a non-synonymous coding single nucleotide polymorphism (SNP) between the B6 and BTBR alleles. Overexpression of the b-tomosyn-2 isoform in the pancreatic β-cell line, INS1 (832/13), resulted in an inhibition of insulin secretion in response to 3 mM 8-bromo cAMP at 7 mM glucose. In vitro binding experiments showed that tomosyn-2 binds recombinant syntaxin-1A and syntaxin-4, key proteins that are involved in insulin secretion via formation of the SNARE complex. The B6 form of tomosyn-2 is more susceptible to proteasomal degradation than the BTBR form, establishing a functional role for the coding SNP in tomosyn-2. We conclude that tomosyn-2 is the major gene responsible for the T2D Chr 16 quantitative trait locus (QTL) we mapped in our mouse cross. Our findings suggest that tomosyn-2 is a key negative regulator of insulin secretion.
Zdroje
1. BillingsLKFlorezJC 2010 The genetics of type 2 diabetes: what have we learned from GWAS? Annals of the New York Academy of Sciences 1212 59 77
2. CleeSMYandellBSSchuelerKMRabagliaMERichardsOC 2006 Positional cloning of Sorcs1, a type 2 diabetes quantitative trait locus. Nat Genet 38 688 693
3. ScherneckSNestlerMVogelHBluherMBlockMD 2009 Positional cloning of zinc finger domain transcription factor Zfp69, a candidate gene for obesity-associated diabetes contributed by mouse locus Nidd/SJL. PLoS Genet 5 e1000541 doi:10.1371/journal.pgen.1000541
4. Dokmanovic-ChouinardMChungWKChevreJCWatsonEYonanJ 2008 Positional cloning of “Lisch-Like”, a candidate modifier of susceptibility to type 2 diabetes in mice. PLoS Genet 4 e1000137 doi:10.1371/journal.pgen.1000137
5. ZhangSHReddickRLPiedrahitaJAMaedaN 1992 Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E. Science 258 468 471
6. ChuaSCJrChungWKWu-PengXSZhangYLiuSM 1996 Phenotypes of mouse diabetes and rat fatty due to mutations in the OB (leptin) receptor. Science 271 994 996
7. HerbergLColemanDL 1977 Laboratory animals exhibiting obesity and diabetes syndromes. Metabolism 26 59 99
8. CleeSMNadlerSTAttieAD 2005 Genetic and genomic studies of the BTBR ob/ob mouse model of type 2 diabetes. Am J Ther 12 491 498
9. StoehrJPNadlerSTSchuelerKLRabagliaMEYandellBS 2000 Genetic obesity unmasks nonlinear interactions between murine type 2 diabetes susceptibility loci. Diabetes 49 1946 1954
10. TrayhurnP 1984 The development of obesity in animals: the role of genetic susceptibility. Clin Endocrinol Metab 13 451 474
11. MiuraAIshizukaTItayaSIshizawaMKanohY 1998 Glucose- and phorbol ester-induced insulin secretion in human insulinoma cells--association with protein kinase C activation. Biochem Mol Biol Int 46 739 745
12. HenquinJC 2000 Triggering and amplifying pathways of regulation of insulin secretion by glucose. Diabetes 49 1751 1760
13. RutterGA 2001 Nutrient-secretion coupling in the pancreatic islet beta-cell: recent advances. Mol Aspects Med 22 247 284
14. LacyPEWalkerMMFinkCJ 1972 Perifusion of isolated rat islets in vitro. Participation of the microtubular system in the biphasic release of insulin. Diabetes 21 987 998
15. ZhangWLiljaLMandicSAGromadaJSmidtK 2006 Tomosyn is expressed in beta-cells and negatively regulates insulin exocytosis. Diabetes 55 574 581
16. KellerMPChoiYWangPDavisDBRabagliaME 2008 A gene expression network model of type 2 diabetes links cell cycle regulation in islets with diabetes susceptibility. Genome research 18 706 716
17. CleeSMAttieAD 2007 The genetic landscape of type 2 diabetes in mice. Endocr Rev 28 48 83
18. ParikhHGroopL 2004 Candidate genes for type 2 diabetes. Rev Endocr Metab Disord 5 151 176
19. NeelJV 1962 Diabetes mellitus: a “thrifty” genotype rendered detrimental by “progress”? Am J Hum Genet 14 353 362
20. BonnycastleLLWillerCJConneelyKNJacksonAUBurrillCP 2006 Common variants in maturity-onset diabetes of the young genes contribute to risk of type 2 diabetes in Finns. Diabetes 55 2534 2540
21. WilliamsALBielopolskiNMerozDLamADPassmoreDR 4553
22. WangZThurmondDC 2009 Mechanisms of biphasic insulin-granule exocytosis - roles of the cytoskeleton, small GTPases and SNARE proteins. J Cell Sci 122 893 903
23. GroffenAJJacobsenLSchutDVerhageM 2005 Two distinct genes drive expression of seven tomosyn isoforms in the mammalian brain, sharing a conserved structure with a unique variable domain. J Neurochem 92 554 568
24. MasudaESHuangBCFisherJMLuoYSchellerRH 1998 Tomosyn binds t-SNARE proteins via a VAMP-like coiled coil. Neuron 21 479 480
25. HatsuzawaKLangTFasshauerDBrunsDJahnR 2003 The R-SNARE motif of tomosyn forms SNARE core complexes with syntaxin 1 and SNAP-25 and down-regulates exocytosis. J Biol Chem 278 31159 31166
26. YamamotoYMochidaSKurookaTSakisakaT 2009 Reciprocal intramolecular interactions of tomosyn control its inhibitory activity on SNARE complex formation. J Biol Chem 284 12480 12490
27. GladychevaSELamADLiuJD'Andrea-MerrinsMYizharO 2007 Receptor-mediated regulation of tomosyn-syntaxin 1A interactions in bovine adrenal chromaffin cells. J Biol Chem 282 22887 22899
28. BabaTSakisakaTMochidaSTakaiY 2005 PKA-catalyzed phosphorylation of tomosyn and its implication in Ca2+-dependent exocytosis of neurotransmitter. J Cell Biol 170 1113 1125
29. McEwenJMMadisonJMDybbsMKaplanJM 2006 Antagonistic regulation of synaptic vesicle priming by Tomosyn and UNC-13. Neuron 51 303 315
30. GrachevaEOBurdinaAOHolgadoAMBerthelot-GrosjeanMAckleyBD 2006 Tomosyn inhibits synaptic vesicle priming in Caenorhabditis elegans. PLoS Biol 4 e261 doi:10.1371/journal.pbio.0040261
31. PobbatiAVRazetoABoddenerMBeckerSFasshauerD 2004 Structural basis for the inhibitory role of tomosyn in exocytosis. J Biol Chem 279 47192 47200
32. YizharOLipsteinNGladychevaSEMattiUErnstSA 2007 Multiple functional domains are involved in tomosyn regulation of exocytosis. J Neurochem 103 604 616
33. YamamotoYMochidaSMiyazakiNKawaiKFujikuraK 0955 Tomosyn inhibits synaptotagmin-1-mediated step of Ca2+-dependent neurotransmitter release through its N-terminal WD40 repeats J Biol Chem 285 40943 40955
34. YamamotoYFujikuraKSakaueMOkimuraKKobayashiY The tail domain of tomosyn controls membrane fusion through tomosyn displacement by VAMP2. Biochem Biophys Res Commun 399 24 30
35. WidbergCHBryantNJGirottiMReaSJamesDE 2003 Tomosyn interacts with the t-SNAREs syntaxin4 and SNAP23 and plays a role in insulin-stimulated GLUT4 translocation. J Biol Chem 278 35093 35101
36. SiddiqiSManiAMSiddiqiSA The identification of the SNARE complex required for the fusion of VLDL-transport vesicle with hepatic cis-Golgi. Biochem J 429 391 401
37. ColemanDLHummelKP 1973 The influence of genetic background on the expression of the obese (Ob) gene in the mouse. Diabetologia 9 287 293
38. RabagliaMEGray-KellerMPFreyBLShortreedMRSmithLM 2005 Alpha-Ketoisocaproate-induced hypersecretion of insulin by islets from diabetes-susceptible mice. Am J Physiol Endocrinol Metab 289 E218 224
39. JewellJLOhEBennettSMMerouehSOThurmondDC 2008 The tyrosine phosphorylation of Munc18c induces a switch in binding specificity from syntaxin 4 to Doc2beta. J Biol Chem 283 21734 21746
40. BhatnagarSDamronHAHillgartnerFB 2009 Fibroblast growth factor-19, a novel factor that inhibits hepatic fatty acid synthesis. J Biol Chem 284 10023 10033
41. MinJOkadaSKanzakiMElmendorfJSCokerKJ 1999 Synip: a novel insulin-regulated syntaxin 4-binding protein mediating GLUT4 translocation in adipocytes. Mol Cell 3 751 760
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Genetika Reprodukčná medicínaČlánok vyšiel v časopise
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