Mutation screening and function prediction of melanocortin-4 receptor gene in obese children

SONG Jie-Yun; WANG Du; MA Jun; WANG Hai-Jun

doi:10.7499/j.issn.1008-8830.2015.04.013

PDF(1372 KB)

Chinese Journal of Contemporary Pediatrics ›› 2015, Vol. 17 ›› Issue (4) : 356-361. DOI: 10.7499/j.issn.1008-8830.2015.04.013

CLINICAL RESEARCH

Mutation screening and function prediction of melanocortin-4 receptor gene in obese children

SONG Jie-Yun, WANG Du, MA Jun, WANG Hai-Jun

Author information +

History +

Abstract

Objective To screen the coding region of melanocortin-4 receptor gene (MC4R) for mutations in children, analyze the association of the identified variants with obesity-related phenotypes, and predict the potential functions of the identified variants. Methods A case-control study was conducted in 160 severely obese children and 100 normal-weight controls, all aged 7-18 years. Their anthropometric data were collected and blood tests were performed. The coding region of MC4R gene was screened by polymerase chain reaction (PCR), single strand conformation polymorphism and sequencing, and the potential functions of the identified variants were predicted by related online databases. Results Three heterozygous missense mutations were identified in obese children (Val95Ile, Val166Ile and Val179Ala), and one heterozygous missense mutation was found in controls (Met218Thr). Val103Ile variant was found to be carried by seven subjects in the obese group and six in the control group (P>0.05). Val179Ala was a newly identified heterozygous mutation. No significant differences in BMI, weight, waist circumstance, hip circumstance, serum lipid parameters, fasting glucose, and body fat percentage were found between Val95Ile, Val166Ile or Val179Ala mutation carriers and non-carriers in obese children. The function prediction of the variants showed that all the five identified variants influenced the protein function. Conclusions Five variants were identified in the coding region of MC4R gene, among which Val179Ala was newly identified. All the five variants might influence the protein function as evidenced by online prediction.

Key words

Melanocortin-4 receptor / Obesity / Gene screening / Function prediction / Child

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

SONG Jie-Yun, WANG Du, MA Jun, WANG Hai-Jun. Mutation screening and function prediction of melanocortin-4 receptor gene in obese children[J]. Chinese Journal of Contemporary Pediatrics. 2015, 17(4): 356-361 https://doi.org/10.7499/j.issn.1008-8830.2015.04.013

References

[1] Tao YX. The melanocortin-4 receptor: physiology, pharmacology, and pathophysiology[J]. Endocr Rev, 2010, 31(4): 506-543.
[2] Fan ZC, Tao YX. Functional characterization and pharmacological rescue of melanocortin-4 receptor mutations identified from obese patients[J]. J Cell Mol Med, 2009, 13(9B): 3268-3282.
[3] Wang ZQ, Tao YX. Functional studies on twenty novel naturally occurring melanocortin-4 receptor mutations[J]. Biochim Biophys Acta, 2011, 1812(9): 1190-1199.
[4] Tao YX. Mutations in melanocortin-4 receptor and human obesity[J]. Prog Mol Biol Transl Sci, 2009, 88: 173-204.
[5] Lee YS. Melanocortin 3 receptor gene and melanocortin 4 receptor gene mutations: the Asian Perspective[J]. Diabetes Metab Res Rev, 2012, 28(Suppl 2): 26-31.
[6] 中国学生体质与健康研究组. 2005年中国学生体质与健康调研报告[M]. 北京: 高等教育出版社, 2007: 193-197.
[7] 中国学生体质与健康研究组. 2005年中国学生体质与健康调研检测细则[M]. 北京: 高等教育出版社, 2007.
[8] Hinney A, Schmidt A, Nottebom K, et al. Several mutations in the melanocortin-4 receptor gene including a nonsense and a frameshift mutation associated with dominantly inherited obesity in humans[J]. J Clin Endocrinol Metab, 1999, 84(4): 1483-1486.
[9] Adzhubei I A, Schmidt S, Peshkin L, et al. A method and server for predicting damaging missense mutations[J]. Nat Methods, 2010, 7(4): 248-249.
[10] Adzhubei I, Jordan DM, Sunyaev SR. Predicting functional effect of human missense mutations using PolyPhen-2[J]. Curr Protoc Hum Genet, 2013, Chapter 7: t7-t20.
[11] Bromberg Y, Yachdav G, Rost B. SNAP predicts effect of mutations on protein function[J]. Bioinformatics, 2008, 24(20): 2397-2398.
[12] Hinney A, Hohmann S, Geller F, et al. Melanocortin-4 receptor gene: case-control study and transmission disequilibrium test confirm that functionally relevant mutations are compatible with a major gene effect for extreme obesity[J]. J Clin Endocrinol Metab, 2003, 88(9): 4258-4267.
[13] Xiang Z, Litherland SA, Sorensen NB, et al. Pharmacological characterization of 40 human melanocortin-4 receptor polymorphisms with the endogenous proopiomelanocortinderived agonists and the agouti-related protein (AGRP) antagonist[J]. Biochemistry, 2006, 45(23): 7277-7288.
[14] Hinney A, Bettecken T, Tarnow P, et al. Prevalence, spectrum, and functional characterization of melanocortin-4 receptor gene mutations in a representative population-based sample and obese adults from Germany[J]. J Clin Endocrinol Metab, 2006, 91(5): 1761-1769.
[15] Rovite V, Petrovska R, Vaivade I, et al. The role of common and rare MC4R variants and FTO polymorphisms in extreme form of obesity[J]. Mol Biol Rep, 2014, 41(3): 1491-1500.
[16] Rong R, Tao YX, Cheung BM, et al. Identification and functional characterization of three novel human melanocortin-4 receptor gene variants in an obese Chinese population[J]. Clin Endocrinol (Oxf), 2006, 65(2): 198-205.
[17] Wang D, Ma J, Zhang S, et al. Association of the MC4R V103I polymorphism with obesity: a Chinese case-control study and meta-analysis in 55,195 individuals[J]. Obesity (Silver Spring), 2010, 18(3): 573-579.
[18] Wangensteen T, Kolsgaard ML, Mattingsdal M, et al. Mutations in the melanocortin 4 receptor (MC4R) gene in obese patients in Norway[J]. Exp Clin Endocrinol Diabetes, 2009, 117(6): 266- 273.
[19] Wang CL, Liang L, Wang HJ, et al. Several mutations in the melanocortin 4 receptor gene are associated with obesity in Chinese children and adolescents[J]. J Endocrinol Invest, 2006, 29(10): 894-898.
[20] 蔡姝冰, 贾伟平, 方启晨, 等. F261S—肥胖患者中黑皮素 4 受体基因的新突变[J]. 中华内分泌代谢杂志, 2004, 20(4): 372-375.
[21] Lee YS, Poh LK, Kek BL, et al. Novel melanocortin 4 receptor gene mutations in severely obese children[J]. Clin Endocrinol (Oxf), 2008, 68(4): 529-535.
[22] 石星, 秦锐, 韩蓓. 南京地区单纯性肥胖患儿黑皮素 4 受体基因筛查[J]. 实用儿科临床杂志, 2010, 25(7): 488-490.
[23] Bromberg Y, Overton J, Vaisse C, et al. In silico mutagenesis: a case study of the melanocortin 4 receptor[J]. FASEB J, 2009, 23(9): 3059-3069.