The Journal of Physiological Sciences

, Volume 69, Issue 1, pp 103–112 | Cite as

Characterization of Δ(G970-T1122)-CFTR, the most frequent CFTR mutant identified in Japanese cystic fibrosis patients

  • Kanako Wakabayashi-Nakao
  • Yingchun Yu
  • Miyuki Nakakuki
  • Tzyh-Chang Hwang
  • Hiroshi Ishiguro
  • Yoshiro SohmaEmail author
Original Paper


A massive deletion over three exons 16-17b in the CFTR gene was identified in Japanese CF patients with the highest frequency (about 70% of Japanese CF patients definitely diagnosed). This pathogenic mutation results in a deletion of 153 amino acids from glycine at position 970 (G970) to threonine at 1122 (T1122) in the CFTR protein without a frameshift. We name it Δ(G970-T1122)-CFTR. In the present study, we characterized the Δ(G970-T1122)-CFTR expressed in CHO cells using immunoblots and a super resolution microscopy. Δ(G970-T1122)-CFTR proteins were synthesized and core-glycosylated but not complex-glycosylated. This observation suggests that the Δ(G970-T1122) mutation can be categorized into the class II mutation like ΔF508. However, VX-809 a CFTR corrector that can help maturation of ΔF508, had no effect on Δ(G970-T1122). Interestingly C-terminal FLAG tag seems to partially rescue the trafficking defect of Δ(G970-T1122)-CFTR; however the rescued Δ(G970-T1122)-CFTR proteins do not assume channel function. Japanese, and perhaps people in other Asian nations, carry a class II mutation Δ(G970-T1122) with a higher frequency than previously appreciated. Further study of the Δ(G970-T1122)-CFTR is essential for understanding CF and CFTR-related diseases particularly in Asian countries.


Cystic fibrosis CFTR Mutation Japanese Asian 



We are grateful to Drs. Yoichiro Abe and Masato Yasui (Keio University) for their useful help and discussions. We are also grateful to the Collaborative Research Resources, Keio University School of Medicine for equipment and technical supports. K.W.-N. is a Japan Society for the Promotion of Sciences (JSPS) Research Fellow. This work was supported by JSPS KAKENHI Grant Numbers 17J40033 (K.W.-N.), 25293049, 15K15035, 16H05122 (Y.S.), and 16K09392 (H.I.).

Author contributions

KW-N designed and performed the molecular biological and biochemical experiments and helped write the manuscript. Y-CY designed and performed the electro-physiological experiments and helped write the manuscript. MN designed the molecular biological experiments and T-CH designed the electro-physiological experiments. HI conceptualized the project and helped write the manuscript. YS conceptualized the project, designed the experiments, and helped write the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    Welsh MJ, Smith AE (1993) Molecular mechanisms of CFTR chloride channel dysfunction in cystic fibrosis. Cell 73(7):1251–1254.
  2. 2.
    Sohma Y, Hwang TC (2015) Cystic fibrosis and the CFTR anion channel. In: Zheng J, Trudeau MC (eds) Handbook of ion channels. CRC Press, Taylor & Francis Books Inc, Oxford, pp 627–648CrossRefGoogle Scholar
  3. 3.
    Zielenski J, Tsui LC (1995) Cystic fibrosis: genotypic and phenotypic variations. Annu Rev Genet 29:777–807. CrossRefGoogle Scholar
  4. 4.
    Singh M, Rebordosa C, Bernholz J, Sharma N (2015) Epidemiology and genetics of cystic fibrosis in Asia: in preparation for the next-generation treatments. Respirology 20(8):1172–1181. CrossRefGoogle Scholar
  5. 5.
    Bosch B, Bilton D, Sosnay P, Raraigh KS, Mak DY, Ishiguro H, Gulmans V, Thomas M, Cuppens H, Amaral M, De Boeck K (2017) Ethnicity impacts the cystic fibrosis diagnosis: a note of caution. J Cyst Fibros. Google Scholar
  6. 6.
    Imaizumi Y (1995) Incidence and mortality rates of cystic fibrosis in Japan, 1969–1992. Am J Med Genet 58(2):161–168. CrossRefGoogle Scholar
  7. 7.
    Yamashiro Y, Shimizu T, Oguchi S, Shioya T, Nagata S, Ohtsuka Y (1997) The estimated incidence of cystic fibrosis in Japan. J Pediatr Gastroenterol Nutr 24(5):544–547.
  8. 8.
    Nakakuki M, Fujiki K, Yamamoto A, Ko SB, Yi L, Ishiguro M, Yamaguchi M, Kondo S, Maruyama S, Yanagimoto K, Naruse S, Ishiguro H (2012) Detection of a large heterozygous deletion and a splicing defect in the CFTR transcripts from nasal swab of a Japanese case of cystic fibrosis. J Hum Genet 57(7):427–433. CrossRefGoogle Scholar
  9. 9.
    Yu YC, Miki H, Nakamura Y, Hanyuda A, Matsuzaki Y, Abe Y, Yasui M, Tanaka K, Hwang TC, Bompadre SG, Sohma Y (2011) Curcumin and genistein additively potentiate G551D-CFTR. J Cyst Fibros 10(4):243–252. CrossRefGoogle Scholar
  10. 10.
    Mio K, Ogura T, Mio M, Shimizu H, Hwang TC, Sato C, Sohma Y (2008) Three-dimensional reconstruction of human cystic fibrosis transmembrane conductance regulator chloride channel revealed an ellipsoidal structure with orifices beneath the putative transmembrane domain. J Biol Chem 283(44):30300–30310. CrossRefGoogle Scholar
  11. 11.
    Shimizu H, Yu YC, Kono K, Kubota T, Yasui M, Li M, Hwang TC, Sohma Y (2010) A stable ATP binding to the nucleotide binding domain is important for reliable gating cycle in an ABC transporter CFTR. J Physiol Sci 60(5):353–362. CrossRefGoogle Scholar
  12. 12.
    Chang XB, Mengos A, Hou YX, Cui L, Jensen TJ, Aleksandrov A, Riordan JR, Gentzsch M (2008) Role of N-linked oligosaccharides in the biosynthetic processing of the cystic fibrosis membrane conductance regulator. J Cell Sci 121(Pt 17):2814–2823. CrossRefGoogle Scholar
  13. 13.
    Riordan JR, Rommens JM, Kerem B, Alon N, Rozmahel R, Grzelczak Z, Zielenski J, Lok S, Plavsic N, Chou JL et al (1989) Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245(4922):1066–1073.
  14. 14.
    Cheng SH, Gregory RJ, Marshall J, Paul S, Souza DW, White GA, O’Riordan CR, Smith AE (1990) Defective intracellular transport and processing of CFTR is the molecular basis of most cystic fibrosis. Cell 63(4):827–834.
  15. 15.
    Lukacs GL, Mohamed A, Kartner N, Chang XB, Riordan JR, Grinstein S (1994) Conformational maturation of CFTR but not its mutant counterpart (∆F508) occurs in the endoplasmic reticulum and requires ATP. EMBO J 13(24):6076–6086.
  16. 16.
    Glozman R, Okiyoneda T, Mulvihill CM, Rini JM, Barriere H, Lukacs GL (2009) N-glycans are direct determinants of CFTR folding and stability in secretory and endocytic membrane traffic. J Cell Biol 184(6):847–862. CrossRefGoogle Scholar
  17. 17.
    Kartner N, Augustinas O, Jensen TJ, Naismith AL, Riordan JR (1992) Mislocalization of ∆F508 CFTR in cystic fibrosis sweat gland. Nat Genet 1(5):321–327. CrossRefGoogle Scholar
  18. 18.
    Van Goor F, Hadida S, Grootenhuis PD, Burton B, Stack JH, Straley KS, Decker CJ, Miller M, McCartney J, Olson ER, Wine JJ, Frizzell RA, Ashlock M, Negulescu PA (2011) Correction of the F508del-CFTR protein processing defect in vitro by the investigational drug VX-809. Proc Natl Acad Sci USA 108(46):18843–18848. CrossRefGoogle Scholar
  19. 19.
    Denning GM, Anderson MP, Amara JF, Marshall J, Smith AE, Welsh MJ (1992) Processing of mutant cystic fibrosis transmembrane conductance regulator is temperature-sensitive. Nature 358(6389):761–764. CrossRefGoogle Scholar
  20. 20.
    He L, Kota P, Aleksandrov AA, Cui L, Jensen T, Dokholyan NV, Riordan JR (2013) Correctors of ∆F508 CFTR restore global conformational maturation without thermally stabilizing the mutant protein. FASEB J 27(2):536–545. CrossRefGoogle Scholar
  21. 21.
    Rubaiy HN, Linsdell P (2015) Location of a permeant anion binding site in the cystic fibrosis transmembrane conductance regulator chloride channel pore. J Physiol Sci 65(3):233–241. CrossRefGoogle Scholar
  22. 22.
    Hwang TC, Yeh JT, Zhang J, Yu YC, Yeh HI, Destefano S (2018) Structural mechanisms of CFTR function and dysfunction. J Gen Physiol 150(4):539–570. Google Scholar
  23. 23.
    Rabeh WM, Bossard F, Xu H, Okiyoneda T, Bagdany M, Mulvihill CM, Du K, di Bernardo S, Liu Y, Konermann L, Roldan A, Lukacs GL (2012) Correction of both NBD1 energetics and domain interface is required to restore ∆F508 CFTR folding and function. Cell 148(1–2):150–163. CrossRefGoogle Scholar
  24. 24.
    Mendoza JL, Schmidt A, Li Q, Nuvaga E, Barrett T, Bridges RJ, Feranchak AP, Brautigam CA, Thomas PJ (2012) Requirements for efficient correction of ∆F508 CFTR revealed by analyses of evolved sequences. Cell 148(1–2):164–174. CrossRefGoogle Scholar
  25. 25.
    Okiyoneda T, Veit G, Dekkers JF, Bagdany M, Soya N, Xu H, Roldan A, Verkman AS, Kurth M, Simon A, Hegedus T, Beekman JM, Lukacs GL (2013) Mechanism-based corrector combination restores ∆F508-CFTR folding and function. Nat Chem Biol 9(7):444–454. CrossRefGoogle Scholar
  26. 26.
    Loo TW, Clarke DM (2017) Corrector VX-809 promotes interactions between cytoplasmic loop one and the first nucleotide-binding domain of CFTR. Biochem Pharmacol. Google Scholar
  27. 27.
    Hall RA, Ostedgaard LS, Premont RT, Blitzer JT, Rahman N, Welsh MJ, Lefkowitz RJ (1998) A C-terminal motif found in the beta2-adrenergic receptor, P2Y1 receptor and cystic fibrosis transmembrane conductance regulator determines binding to the Na+/H+ exchanger regulatory factor family of PDZ proteins. Proc Natl Acad Sci USA 95(15):8496–8501.
  28. 28.
    Short DB, Trotter KW, Reczek D, Kreda SM, Bretscher A, Boucher RC, Stutts MJ, Milgram SL (1998) An apical PDZ protein anchors the cystic fibrosis transmembrane conductance regulator to the cytoskeleton. J Biol Chem 273(31):19797–19801.
  29. 29.
    Cheng J, Moyer BD, Milewski M, Loffing J, Ikeda M, Mickle JE, Cutting GR, Li M, Stanton BA, Guggino WB (2002) A Golgi-associated PDZ domain protein modulates cystic fibrosis transmembrane regulator plasma membrane expression. J Biol Chem 277(5):3520–3529. CrossRefGoogle Scholar
  30. 30.
    Arora K, Moon C, Zhang W, Yarlagadda S, Penmatsa H, Ren A, Sinha C, Naren AP (2014) Stabilizing rescued surface-localized ∆f508 CFTR by potentiation of its interaction with Na(+)/H(+) exchanger regulatory factor 1. Biochemistry 53(25):4169–4179. CrossRefGoogle Scholar
  31. 31.
    Loureiro CA, Matos AM, Dias-Alves A, Pereira JF, Uliyakina I, Barros P, Amaral MD, Matos P (2015) A molecular switch in the scaffold NHERF1 enables misfolded CFTR to evade the peripheral quality control checkpoint. Sci Signal 8(377):ra48. CrossRefGoogle Scholar
  32. 32.
    Wang S, Yue H, Derin RB, Guggino WB, Li M (2000) Accessory protein facilitated CFTR-CFTR interaction, a molecular mechanism to potentiate the chloride channel activity. Cell 103(1):169–179.
  33. 33.
    Raghuram V, Mak DO, Foskett JK (2001) Regulation of cystic fibrosis transmembrane conductance regulator single-channel gating by bivalent PDZ-domain-mediated interaction. Proc Natl Acad Sci USA 98(3):1300–1305. CrossRefGoogle Scholar
  34. 34.
    Benharouga M, Sharma M, So J, Haardt M, Drzymala L, Popov M, Schwapach B, Grinstein S, Du K, Lukacs GL (2003) The role of the C terminus and Na+/H+ exchanger regulatory factor in the functional expression of cystic fibrosis transmembrane conductance regulator in nonpolarized cells and epithelia. J Biol Chem 278(24):22079–22089. CrossRefGoogle Scholar
  35. 35.
    Cheng J, Wang H, Guggino WB (2004) Modulation of mature cystic fibrosis transmembrane regulator protein by the PDZ domain protein CAL. J Biol Chem 279(3):1892–1898. CrossRefGoogle Scholar
  36. 36.
    Sharma N, LaRusch J, Sosnay PR, Gottschalk LB, Lopez AP, Pellicore MJ, Evans T, Davis E, Atalar M, Na CH, Rosson GD, Belchis D, Milewski M, Pandey A, Cutting GR (2016) A sequence upstream of canonical PDZ-binding motif within CFTR COOH-terminus enhances NHERF1 interaction. Am J Physiol Lung Cell Mol Physiol 311(6):L1170–L1182. CrossRefGoogle Scholar
  37. 37.
    Benharouga M, Haardt M, Kartner N, Lukacs GL (2001) COOH-terminal truncations promote proteasome-dependent degradation of mature cystic fibrosis transmembrane conductance regulator from post-Golgi compartments. J Cell Biol 153(5):957–970.
  38. 38.
    McClure ML, Barnes S, Brodsky JL, Sorscher EJ (2016) Trafficking and function of the cystic fibrosis transmembrane conductance regulator: a complex network of posttranslational modifications. Am J Physiol Lung Cell Mol Physiol 311(4):L719–L733. CrossRefGoogle Scholar
  39. 39.
    Castellani C, Cuppens H, Macek M Jr, Cassiman JJ, Kerem E, Durie P, Tullis E, Assael BM, Bombieri C, Brown A, Casals T, Claustres M, Cutting GR, Dequeker E, Dodge J, Doull I, Farrell P, Ferec C, Girodon E, Johannesson M, Kerem B, Knowles M, Munck A, Pignatti PF, Radojkovic D, Rizzotti P, Schwarz M, Stuhrmann M, Tzetis M, Zielenski J, Elborn JS (2008) Consensus on the use and interpretation of cystic fibrosis mutation analysis in clinical practice. J Cyst Fibros 7(3):179–196. CrossRefGoogle Scholar
  40. 40.
    Girardet A, Guittard C, Altieri JP, Templin C, Stremler N, Beroud C, Des Georges M, Claustres M (2007) Negative genetic neonatal screening for cystic fibrosis caused by compound heterozygosity for two large CFTR rearrangements. Clin Genet 72(4):374–377. CrossRefGoogle Scholar

Copyright information

© The Physiological Society of Japan and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmaceutical Sciences and Center for Medical SciencesInternational University of Health and WelfareOtawaraJapan
  2. 2.Dalton Cardiovascular Research CenterUniversity of MissouriColumbiaUSA
  3. 3.Department of Medical Pharmacology and PhysiologyUniversity of MissouriColumbiaUSA
  4. 4.Department of Human NutritionNagoya University Graduate School of MedicineNagoyaJapan
  5. 5.Department of PharmacologyKeio University School of MedicineTokyoJapan

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