Action of the dominant-negative effect on the pathogenesis of insulinopathies_West China Medical Journal

Authors：

 SUN Nan ¹ , HAN Li ¹ , Sayila·Haimiti ¹ , CUI Jingqiu ² , WANG Xinling ¹

1. Department of Endocrinology, Xinjiang Uygur Autonomous Region People’s Hospital, Urumqi, Xinjiang 830001, P. R. China;
2. Department of Endocrinology, the General Hospital of Tianjin Medical University, Tianjin 300052, P. R. China;

Corresponding author：

SUN Nan, Email: sunnan07@163.com

Keywords：

Diabetes; Dominant-negative effect; Insulin; Gene mutation

DOI：

10.7507/1002-0179.201809080

Video：

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Abstract Full text Figures/Tables Video References Cited by

ObjectiveTo explore the action of dominant-negative effect on mutant insulin gene-induced diabetes.Methods293T cells were transfected with a recombinant plasmid containing mutant preproinsulinogen complementary DNA (cDNA) and a recombinant plasmid containing human wild-type preproinsulinogen cDNA. There were 5 mutant groups which mutant preproinsulins respectively bear substitutions V(A3)L, C(A7)Y, R(SP6)H, G(B8)S or G(C28)R. Wild-type mouse preproinsulin and wild-type human preproinsulin were co-transfected as normal control group. After 48 hours, medium and cells were collected. Human proinsulin were detected by human-specific proinsulin radioimmunoassay.ResultsCompared with the control group [(135.84±1.89) pmol/L], human proinsulin levels in medium of C(A7)Y group [(29.28±6.85) pmol/L] and G(B8)S group[(33.62±10.52) pmol/L] decreased significantly (P<0.01). There was no significant difference in human proinsulin level between the other groups and the control group (P>0.05).ConclusionMutants C(A7)Y and G(B8)S induce the dominant-negative effect on co-existing wild-type proinsulin.

Citation： SUN Nan, HAN Li, Sayila·Haimiti, CUI Jingqiu, WANG Xinling. Action of the dominant-negative effect on the pathogenesis of insulinopathies. West China Medical Journal, 2018, 33(11): 1406-1410. doi: 10.7507/1002-0179.201809080 Copy

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2.	Støy J, Edghill EL, Flanagan SE, et al. Insulin gene mutations as a cause of permanent neonatal diabetes. Proc Natl Acad Sci USA, 2007, 104(38): 15040-15044.
3.	Bonfanti R, Colombo C, Nocerino V, et al. Insulin gene mutations as cause of diabetes in children negative for five type 1 diabetes autoantibodies. Diabetes Care, 2009, 32(1): 123-125.
4.	Meur G, Simon A, Harun N, et al. Insulin gene mutations resulting in early-onset diabetes: marked differences in clinical presentation, metabolic status, and pathogenic effect through endoplasmic reticulum retention. Diabetes, 2010, 59(3): 653-661.
5.	Hodish I, Liu M, Rajpal G, et al. Misfolded proinsulin affects bystander proinsulin in neonatal diabetes. J Biol Chem, 2010, 285(1): 685-694.
6.	Park SY, Ye H, Steiner DF, et al. Mutant proinsulin proteins associated with neonatal diabetes are retained in the endoplasmic reticulum and not efficiently secreted. Biochem Biophys Res Commun, 2010, 391(3): 1449-1454.
7.	Steiner DF, Tager HS, Chan SJ, et al. Lessons learned from molecular biology of insulin-gene mutations. Diabetes Care, 1990, 13(6): 600-609.
8.	Colombo C, Porzio O, Liu M, et al. Seven mutations in the human insulin gene linked to permanent neonatal/infancy-onset diabetes mellitus. J Clin Invest, 2008, 118(6): 2148-2156.
9.	Haataja L, Manickam N, Soliman A, et al. Disulfide mispairing during proinsulin folding in the endoplasmic reticulum. Diabetes, 2016, 65(4): 1050-1060.
10.	Kim YH, Kastner K, Abdul-Wahid B, et al. Evaluation of conformational changes in diabetes-associated mutation in insulin a chain: a molecular dynamics study. Proteins, 2015, 83(4): 662-669.
11.	Renner S, Braun-Reichhart C, Blutke A, et al. Permanent neonatal diabetes in INS(C94Y) transgenic pigs. Diabetes, 2013, 62(5): 1505-1511.
12.	Blutke A, Renner S, Flenkenthaler F, et al. The Munich MIDY pig biobank: a unique resource for studying organ crosstalk in diabetes. Mol Metab, 2017, 6(8): 931-940.
13.	Støy J, Olsen J, Park SY, et al. In vivo measurement and biological characterisation of the diabetes-associated mutant insulin p.R46Q (GlnB22-insulin). Diabetologia, 2017, 60(8): 1423-1431.
14.	Haataja L, Snapp E, Wright J, et al. Proinsulin intermolecular interactions during secretory trafficking in pancreatic β cells. J Biol Chem, 2013, 288(3): 1896-1906.
15.	Wright J, Birk J, Haataja L, et al. Endoplasmic reticulum oxidoreductin-1α (Ero1α) improves folding and secretion of mutant proinsulin and limits mutant proinsulin-induced endoplasmic reticulum stress. J Biol Chem, 2013, 288(43): 31010-31018.
16.	He K, Cunningham CN, Manickam N, et al. PDI reductase acts on Akita mutant proinsulin to initiate retrotranslocation along the Hrd1/Sel1L-p97 axis. Mol Biol Cell, 2015, 26(19): 3413-3423.
17.	Cunningham CN, He K, Arunagiri A, et al. Chaperone-driven degradation of a misfolded proinsulin mutant in parallel with restoration of wild-type insulin secretion. Diabetes, 2017, 66(3): 741-753.

1. Guo H, Xiong Y, Witkowski P, et al. Inefficient translocation of preproinsulin contributes to pancreatic β cell failure and late-onset diabetes. J Biol Chem, 2014, 289(23): 16290-16302.
2. Støy J, Edghill EL, Flanagan SE, et al. Insulin gene mutations as a cause of permanent neonatal diabetes. Proc Natl Acad Sci USA, 2007, 104(38): 15040-15044.
3. Bonfanti R, Colombo C, Nocerino V, et al. Insulin gene mutations as cause of diabetes in children negative for five type 1 diabetes autoantibodies. Diabetes Care, 2009, 32(1): 123-125.
4. Meur G, Simon A, Harun N, et al. Insulin gene mutations resulting in early-onset diabetes: marked differences in clinical presentation, metabolic status, and pathogenic effect through endoplasmic reticulum retention. Diabetes, 2010, 59(3): 653-661.
5. Hodish I, Liu M, Rajpal G, et al. Misfolded proinsulin affects bystander proinsulin in neonatal diabetes. J Biol Chem, 2010, 285(1): 685-694.
6. Park SY, Ye H, Steiner DF, et al. Mutant proinsulin proteins associated with neonatal diabetes are retained in the endoplasmic reticulum and not efficiently secreted. Biochem Biophys Res Commun, 2010, 391(3): 1449-1454.
7. Steiner DF, Tager HS, Chan SJ, et al. Lessons learned from molecular biology of insulin-gene mutations. Diabetes Care, 1990, 13(6): 600-609.
8. Colombo C, Porzio O, Liu M, et al. Seven mutations in the human insulin gene linked to permanent neonatal/infancy-onset diabetes mellitus. J Clin Invest, 2008, 118(6): 2148-2156.
9. Haataja L, Manickam N, Soliman A, et al. Disulfide mispairing during proinsulin folding in the endoplasmic reticulum. Diabetes, 2016, 65(4): 1050-1060.
10. Kim YH, Kastner K, Abdul-Wahid B, et al. Evaluation of conformational changes in diabetes-associated mutation in insulin a chain: a molecular dynamics study. Proteins, 2015, 83(4): 662-669.
11. Renner S, Braun-Reichhart C, Blutke A, et al. Permanent neonatal diabetes in INS(C94Y) transgenic pigs. Diabetes, 2013, 62(5): 1505-1511.
12. Blutke A, Renner S, Flenkenthaler F, et al. The Munich MIDY pig biobank: a unique resource for studying organ crosstalk in diabetes. Mol Metab, 2017, 6(8): 931-940.
13. Støy J, Olsen J, Park SY, et al. In vivo measurement and biological characterisation of the diabetes-associated mutant insulin p.R46Q (GlnB22-insulin). Diabetologia, 2017, 60(8): 1423-1431.
14. Haataja L, Snapp E, Wright J, et al. Proinsulin intermolecular interactions during secretory trafficking in pancreatic β cells. J Biol Chem, 2013, 288(3): 1896-1906.
15. Wright J, Birk J, Haataja L, et al. Endoplasmic reticulum oxidoreductin-1α (Ero1α) improves folding and secretion of mutant proinsulin and limits mutant proinsulin-induced endoplasmic reticulum stress. J Biol Chem, 2013, 288(43): 31010-31018.
16. He K, Cunningham CN, Manickam N, et al. PDI reductase acts on Akita mutant proinsulin to initiate retrotranslocation along the Hrd1/Sel1L-p97 axis. Mol Biol Cell, 2015, 26(19): 3413-3423.
17. Cunningham CN, He K, Arunagiri A, et al. Chaperone-driven degradation of a misfolded proinsulin mutant in parallel with restoration of wild-type insulin secretion. Diabetes, 2017, 66(3): 741-753.

West China Medical Journal

Action of the dominant-negative effect on the pathogenesis of insulinopathies

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