Skip to main content
SpringerLink
Log in

A novel Cx50 (GJA8) p.H277Y mutation associated with autosomal dominant congenital cataract identified with targeted next-generation sequencing

  • Genetics
  • Published:
Graefe's Archive for Clinical and Experimental Ophthalmology Aims and scope Submit manuscript

Abstract

Background

To unravel the molecular genetic background responsible for autosomal dominant congenital pulverulent nuclear cataracts in a four-generation Chinese family.

Methods

Family history data were collected, ophthalmological examinations were performed, and genomic DNA was extracted from peripheral blood of the family members. The candidate genes were captured and sequenced by targeted next-generation sequencing, and the results were confirmed by Sanger sequencing. The structure modelling of the protein was displayed based on Swiss-Model Server, and its possible changes in the secondary structure were predicted using Antheprot 2000 software. The chemical dissimilarity and possible functional impact of an amino acid substitution were performed with Grantham score, PolyPhen-2, and SIFT predictions. Protein distributions were assessed by confocal microscopy.

Results

A novel heterozygous c.829C > T transition that led to the substitution of a highly conserved histidine by tyrosine at codon 277 (p.H277Y) in the coding region of connexin50 (Cx50, GJA8) was identified. Bioinformatics analysis showed that the mutation likely altered the secondary structure of the protein by replacing the helix of the COOH-terminal portion with a turn. The mutation was predicted to be moderately conservative by Grantham score and to be deleterious by both PolyPhen-2 and SIFT with consistent results. In addition, when expressed in COS1 cells, the mutation led to protein accumulation and caused changes in Cx 50 protein localization pattern.

Conclusions

This is a novel missense mutation [c.829C > T, (p.H277Y)] identified in exon 2 of Cx50. Our findings expand the spectrum of Cx50 mutations that are associated with autosomal dominant congenital pulverulent nuclear cataract.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Wang KJ, Wang S, Cao NQ, Yan YB, Zhu SQ (2011) A novel mutation in CRYBB1 associated with congenital cataract-microcornea syndrome: the p.Ser129Arg mutation destabilizes the βB1/βA3-crystallin heteromer but not the βB1-crystallin homomer. Hum Mutat 32:E2050–E2060

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Reddy MA, Francis PJ, Berry V, Bhattacharya SS, Moore AT (2004) Molecular genetic basis of inherited cataract and associated phenotypes. Surv Ophthalmol 49:300–315

    Article  PubMed  Google Scholar 

  3. Holmes JM, Leske DA, Burke JP, Hodge DO (2003) Birth prevalence of visually significant infantile cataract in a defined U.S. population. Ophthalmic Epidemiol 10:67–74

    Article  PubMed  Google Scholar 

  4. Apple DJ, Ram J, Foster A, Peng Q (2000) Elimination of cataract blindness: a global perspective entering the new millennium. Surv Ophthalmol 45(Suppl 1):S1–S196

    PubMed  Google Scholar 

  5. Gao X, Cheng J, Lu C, Li X, Li F, Liu C, Zhang M, Zhu S, Ma X (2010) A novel mutation in the connexin 50 gene (GJA8) associated with autosomal dominant congenital nuclear cataract in a Chinese family. Curr Eye Res 35:597–604

    Article  CAS  PubMed  Google Scholar 

  6. Hu S, Wang B, Zhou Z, Zhou G, Wang J, Ma X, Qi Y (2010) A novel mutation in GJA8 causing congenital cataract-microcornea syndrome in a Chinese pedigree. Mol Vis 16:1585–1592

    PubMed Central  CAS  PubMed  Google Scholar 

  7. Haargaard B, Wohlfahrt J, Fledelius HC, Rosenberg T, Melbye M (2004) Incidence and cumulative risk of childhood cataract in a cohort of 2.6 million Danish children. Invest Ophthalmol Vis Sci 45:1316–1320

    Article  PubMed  Google Scholar 

  8. Wang K, Wang B, Wang J, Zhou S, Yun B, Suo P, Cheng J, Ma X, Zhu S (2009) A novel GJA8 mutation (p.I31T) causing autosomal dominant congenital cataract in a Chinese family. Mol Vis 15:2813–2820

    PubMed Central  CAS  PubMed  Google Scholar 

  9. Hejtmancik JF (2008) Congenital cataracts and their molecular genetics. Semin Cell Dev Biol 19:134–149

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Arora A, Minogue PJ, Liu X, Reddy MA, Ainsworth JR, Bhattacharya SS, Webster AR, Hunt DM, Ebihara L, Moore AT, Beyer EC, Berthoud VM (2006) A novel GJA8 mutation is associated with autosomal dominant lamellar pulverulent cataract: further evidence for gap junction dysfunction in human cataract. J Med Genet 43:e2

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Mackay DS, Bennett TM, Culican SM, Shiels A (2014) Exome sequencing identifies novel and recurrent mutations in GJA8 and CRYGD associated with inherited cataract. Hum Genomics 8:19

    Article  PubMed Central  PubMed  Google Scholar 

  12. Santana A, Waiswo M (2011) The genetic and molecular basis of congenital cataract. Arq Bras Oftalmol 74:136–142

    Article  PubMed  Google Scholar 

  13. Shiels A, Bennett TM, Knopf HL, Yamada K, Yoshiura K, Niikawa N, Shim S, Hanson PI (2007) CHMP4B, a novel gene for autosomal dominant cataracts linked to chromosome 20q. Am J Hum Genet 81:596–606

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Shiels A, Bennett TM, Knopf HL, Maraini G, Li A, Jiao X, Hejtmancik JF (2008) The EPHA2 gene is associated with cataracts linked to chromosome 1p. Mol Vis 14:2042–2055

    PubMed Central  CAS  PubMed  Google Scholar 

  15. Kaul H, Riazuddin SA, Shahid M, Kousar S, Butt NH, Zafar AU, Khan SN, Husnain T, Akram J, Hejtmancik JF, Riazuddin S (2010) Autosomal recessive congenital cataract linked to EPHA2 in a consanguineous Pakistani family. Mol Vis 16:511–517

    PubMed Central  CAS  PubMed  Google Scholar 

  16. Ge XL, Zhang Y, Wu Y, Lv J, Zhang W, Jin ZB, Qu J, Gu F (2014) Identification of a novel GJA8 (C×50) point mutation causes human dominant congenital cataracts. Sci Rep 4:4121

    PubMed Central  PubMed  Google Scholar 

  17. He Y, Wu J, Dressman DC, Iacobuzio-Donahue C, Markowitz SD, Velculescu VE, Diaz LA Jr, Kinzler KW, Vogelstein B, Papadopoulos N (2010) Heteroplasmic mitochondrial DNA mutations in normal and tumour cells. Nature 464:610–614

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Zenteno JC, Crespí J, Buentello-Volante B, Buil JA, Bassaganyas F, Vela-Segarra JI, Diaz-Cascajosa J, Marieges MT (2014) Next generation sequencing uncovers a missense mutation in COL4A1 as the cause of familial retinal arteriolar tortuosity. Graefes Arch Clin Exp Ophthalmol 252:1789–1794

    Article  CAS  PubMed  Google Scholar 

  19. Yang Y, Wu J, Liu H, Chen X, Wang Y, Zhao M, He X (2013) Two homozygous nonsense mutations of GNPTAB gene in two Chinese families with mucolipidosis II alpha/beta using targeted next-generation sequencing. Genomics 102:169–173

    Article  CAS  PubMed  Google Scholar 

  20. Arnold K, Bordoli L, Kopp J, Schwede T (2006) The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22:195–201

    Article  CAS  PubMed  Google Scholar 

  21. Grantham R (1974) Amino acid difference formula to help explain protein evolution. Science 185:862–864

    Article  CAS  PubMed  Google Scholar 

  22. Li WH, Wu CI, Luo CC (1984) Nonrandomness of point mutation as reflected in nucleotide substitutions in pseudogenes and its evolutionary implications. J Mol Evol 21:58–71

    Article  CAS  PubMed  Google Scholar 

  23. Maeda S, Nakagawa S, Suga M, Yamashita E, Oshima A, Fujiyoshi Y, Tsukihara T (2009) Structure of the connexin 26 gap junction channel at 3.5 a resolution. Nature 458:597–602

    Article  CAS  PubMed  Google Scholar 

  24. Xin L, Gong XQ, Bai D (2010) The role of amino terminus of mouse C×50 in determining transjunctional voltage-dependent gating and unitary conductance. Biophys J 99:2077–2086

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  25. Beyer EC, Berthoud VM (2014) Connexin hemichannels in the lens. Front Physiol 5:20

    Article  PubMed Central  PubMed  Google Scholar 

  26. Yeager M, Harris AL (2007) Gap junction channel structure in the early 21st century: Facts and fantasies. Curr Opin Cell Biol 19:521–528

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Wang N, De Bock M, Decrock E, Bol M, Gadicherla A, Vinken M, Rogiers V, Bukauskas FF, Bultynck G, Leybaert L (2013) Paracrine signaling through plasma membrane hemichannels. Biochim Biophys Acta 1828:35–50

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Sellitto C, Li L, White TW (2004) Connexin50 is essential for normal postnatal lens cell proliferation. Invest Ophthalmol Vis Sci 45:3196–3202

    Article  PubMed  Google Scholar 

  29. Pal JD, Berthoud VM, Beyer EC, Mackay D, Shiels A, Ebihara L (1999) Molecular mechanism underlying a C×50-linked congenital cataract. Am J Physiol 276:C1443–C1446

    CAS  PubMed  Google Scholar 

  30. Somaraju Chalasani ML, Muppirala MG, Ponnam SP, Kannabiran C, Swarup G (2012) A cataract-causing connexin 50 mutant is mislocalized to the ER due to loss of the fourth transmembrane domain and cytoplasmic domain. FEBS Open Bio 3:22–29

    Article  PubMed Central  PubMed  Google Scholar 

  31. Minogue PJ, Tong JJ, Arora A, Russell-Eggitt I, Hunt DM, Moore AT, Ebihara L, Beyer EC, Berthoud VM (2009) A mutant connexin50 with enhanced hemichannel function leads to cell death. Invest Ophthalmol Vis Sci 50:5837–5845

    Article  PubMed Central  PubMed  Google Scholar 

  32. Polyakov A, Shagina I, Khlebnikova O, Evgrafov O (2001) Mutation in the connexin 50 gene (GJA8) in a Russian family with zonular pulverulent cataract. Clin Genet 60:476–478

    Article  CAS  PubMed  Google Scholar 

  33. Hansen L, Mikkelsen A, Nürnberg P, Nürnberg G, Anjum I, Eiberg H, Rosenberg T (2009) Comprehensive mutational screening in a cohort of Danish families with hereditary congenital cataract. Invest Ophthalmol Vis Sci 50:3291–3303

    Article  PubMed  Google Scholar 

  34. Yan M, Xiong CL, Ye SQ, Chen YM, Ke M, Zheng F, Zhou X (2008) A novel connexin 50 (GJA8) mutation in a Chinese family with a dominant congenital pulverulent nuclear cataract. Mol Vis 14:418–424

    PubMed Central  CAS  PubMed  Google Scholar 

  35. Kumar M, Agarwal T, Khokhar S, Kumar M, Kaur P, Roy TS, Dada R (2011) Mutation screening and genotype phenotype correlation of α-crystallin, γ-crystallin and GJA8 gene in congenital cataract. Mol Vis 17:693–707

    PubMed Central  CAS  PubMed  Google Scholar 

  36. Bouvier D, Spagnol G, Chenavas S, Kieken F, Vitrac H, Brownell S, Kellezi A, Forge V, Sorgen PL (2009) Characterization of the structure and intermolecular interactions between the connexin40 and connexin43 carboxyl-terminal and cytoplasmic loop domains. J Biol Chem 284:34257–34271

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Yan N, Zhao Y, Wang Y, Xie A, Huang H, Yu W, Liu X, Cai SP (2011) Molecular genetics of familial nystagmus complicated with cataract and iris anomalies. Mol Vis 17:2612–2617

    PubMed Central  CAS  PubMed  Google Scholar 

  38. Lichtenstein A, Gaietta GM, Deerinck TJ, Crum J, Sosinsky GE, Beyer EC, Berthoud VM (2009) The cytoplasmic accumulations of the cataract-associated mutant, Connexin50P88S, are long-lived and form in the endoplasmic reticulum. Exp Eye Res 88:600–609

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  39. Lichtenstein A, Minogue PJ, Beyer EC, Berthoud VM (2011) Autophagy: a pathway that contributes to connexin degradation. J Cell Sci 124:910–920

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  40. Vanita V, Singh JR, Singh D, Varon R, Sperling K (2008) A mutation in GJA8 (p.P88Q) is associated with “balloon-like” cataract with Y-sutural opacities in a family of Indian origin. Mol Vis 14:1171–1175

    PubMed Central  CAS  PubMed  Google Scholar 

  41. Beyer EC, Ebihara L, Berthoud VM (2013) Connexin mutants and cataracts. Front Pharmacol 4:43

    PubMed Central  CAS  PubMed  Google Scholar 

  42. Su D, Yang Z, Li Q, Guan L, Zhang HED, Zhang L, Zhu S, Ma X (2013) Identification and functional analysis of GJA8 mutation in a Chinese family with autosomal dominant perinuclear cataracts. PLoS One 8:e59926

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  43. Zhu Y, Yu H, Wang W, Gong X, Yao K (2014) A Novel GJA8 Mutation (p.V44A) Causing Autosomal Dominant Congenital Cataract. PLoS One 9:e115406

    Article  PubMed Central  PubMed  Google Scholar 

  44. Rubinos C, Villone K, Mhaske PV, White TW, Srinivas M (2014) Functional effects of C × 50 mutations associated with congenital cataracts. Am J Physiol Cell Physiol 306:C212–C220

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful for all the participating members of the congenital cataract family. We also express our sincere gratitude to Daoyun Zhang, Yunbing Zhang, Chencheng Wang, Zhongqiong Lu, and Qingfu Wang for their technical assistance. This work was supported by grants from the National Science and Technology Pillar Program of the Twelfth Five-year Plan (2011ZX09302-007-02) and the Research Fund for the National Nature Science Funding of China (No. 81273424 and 81170862).

Conflict of interest

All authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest, or non-financial interest in the subject matter or materials discussed in this manuscript.

Author information

Authors and Affiliations

  1. Department of Ophthalmology, Shanghai First People’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

    Chong Chen, Qiao Sun, Kun Liu, Yong Sun & Xun Xu

  2. Shanghai Key Laboratory of Fundus Disease, Shanghai, China

    Chong Chen, Qiao Sun, Kun Liu, Yong Sun & Xun Xu

  3. Department of Medical Genetics, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

    Mingmin Gu

Authors
  1. Chong Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qiao Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mingmin Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kun Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yong Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xun Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yong Sun.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 163 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, C., Sun, Q., Gu, M. et al. A novel Cx50 (GJA8) p.H277Y mutation associated with autosomal dominant congenital cataract identified with targeted next-generation sequencing. Graefes Arch Clin Exp Ophthalmol 253, 915–924 (2015). https://doi.org/10.1007/s00417-015-3019-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00417-015-3019-x

Keywords

Search

Navigation