Carbonic anhydrase II deficiency syndrome is an autosomal recessive osteopetrosis with renal tubular acidosis and cerebral calcifications. We tried to detect the causative mutation for carbonic anhydrase II deficiency syndrome in a five-generation Chinese family.
Materials and Methods
Genomic DNA was extracted from whole blood of the proband, his grandmother, parents, aunt, uncle and sister. The exomes were sequenced by whole exon sequencing followed by genetic analysis and Sanger sequencing validation. Then, physical and chemical properties studies and structure analysis were performed on mutated protein. Finally, Minigene model of vector plasmids for wild type and mutant type was constructed and transfected into human embryonic kidney 293T cells to further explore the expression change of CA2 transcript and protein after mutation.
Sequencing and genetic analysis have revealed the homozygous nonsense mutation of CA2 gene (c.368G > A, p.W123X) in the exon 4 of chromosome 8 of the proband, while it was not found in his grandmother, parents, aunt, uncle and sister. Furthermore, Sanger sequencing in the proband and his parents validated the mutation. Properties and structure of mutated CA2 proteins changed after mutation, especially in change of protein modification and hindrance of zinc ions binding, which may lead to decreased protein expression level of CA2.
We found a new homozygous nonsense mutation in CA2 gene (c.368G > A, p.W123X), which may be valuable in the early diagnosis and therapy of carbonic anhydrase II deficiency syndrome.
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Adamus G, Karren L (2009) Autoimmunity against carbonic anhydrase II affects retinal cell functions in autoimmune retinopathy. J Autoimmun 32:0–139
Al-Ibrahim A, Al-Harbi M, Al-Musallam S (2003) Paralysis episodes in carbonic anhydrase II deficiency. Saudi J Kidney Dis Transpl 14:70–74
Aronow ME, Adamus G, Abu-Asab M, Wang Y, Chan C-C, Zakov ZN, Singh AD (2012) Paraneoplastic vitelliform retinopathy: clinicopathologic correlation review of the literature. Surv Ophthalmol 57:558–564
Baralle D, Baralle M (2005) Splicing in action: assessing disease causing sequence changes. J Med Genet 42:737–748. https://doi.org/10.1136/jmg.2004.029538
Berg JM, Tymoczko JL, Stryer L (2002) Making a fast reaction faster: carbonic anhydrases. W H Freeman, New York
Borthwick JK (2003) A phenocopy of CAII deficiency: a novel genetic explanation for inherited infantile osteopetrosis with distal renal tubular acidosis. J Med Genet 40:115–121
Boulpaep E, Boron W (2009) Medical physiology: a cellular and molecular approach. Saunders/Elsevier, Philadelphia, PA
Cooper TA (2005) Use of minigene systems to dissect alternative splicing elements. Methods 37:0–340
D’Adamo MC et al (2016) A calsequestrin-1 mutation associated with a skeletal muscle disease alters sarcoplasmic Ca2 + release. PLoS One 11:e0155516. https://doi.org/10.1371/journal.pone.0155516
David JP, Rincon M, Neff L, Horne WC, Baron R (2001) Carbonic anhydrase II is an AP-1 target gene in osteoclasts. J Cell Physiol 188(1):89–97
Fujisaki K et al (2007) Receptor activator of NF-κB ligand induces the expression of carbonic anhydrase II, cathepsin K, and matrix metalloproteinase-9 in osteoclast precursor RAW264.7 cells. Life Sci 80:0–1318
Gaildrat P, Killian A, Martins A, Tournier I, Tosi M (2010) Use of splicing reporter minigene assay to evaluate the effect on splicing of unclassified genetic variants. Methods Mol Biol 653:249–57
Hu PY, Roth DE, Skaggs LA, Venta PJ, Tashian RE, Guibaud P, Sly WS (1992) A splice junction mutation in intron 2 of the carbonic anhydrase II gene of osteopetrosis patients from Arabic countries. Hum Mutat 1:288–292
Hu PY, Lim EJ, Ciccolella J, Strisciuglio P, Sly WS (1997) Seven novel mutations in carbonic anhydrase II deficiency syndrome identified by SSCP and direct sequencing analysis. Hum Mutat 9:383–387
Kumpulainen T (1984) Immunohistochemical localization of human carbonic anhydrase isozymes. Ann N Y Acad Sci 429:359–68
Lin SH, Yang SS, Chau T (2010) A practical approach to genetic hypokalemia . Electrolyte Blood Press 8:38–50. https://doi.org/10.5049/ebp.2010.8.1.38
Lindskog S (1997) Structure and mechanism of carbonic anhydrase. Pharmacol Ther 74:1–20. https://doi.org/10.1016/s0163-7258(96)00198-2
Liu Y et al (2017) CPVT-associated cardiac ryanodine receptor mutation G357S with reduced penetrance impairs Ca2 + release termination and diminishes protein expression. PLoS One 12:e0184177. https://doi.org/10.1371/journal.pone.0184177
McKenna R, Frost SC (2014) Overview of the carbonic anhydrase family. Sub Cell Biochem 75:3–5. https://doi.org/10.1007/978-94-007-7359-2_1
Muzalef A, Alshehri M, Al-Abidi A, Al-Trabolsi HA (2005) Marble brain disease in two Saudi Arabian siblings . Ann Trop Paediatr 25:213–218
Pang Q, Qi X, Jiang Y, Wang O, Li M, Xing X, Dong J, Xia W (2015) Two novel CAII mutations causing carbonic anhydrase II deficiency syndrome in two unrelated Chinese families. Metab Brain Dis 30:989–997
Pollard A, Shephard F, Freed J, Liddell S, Chakrabarti L (2016) Mitochondrial proteomic profiling reveals increased carbonic anhydrase II in aging neurodegeneration. Aging (Albany NY) 8:2425–2436. https://doi.org/10.18632/aging.101064
Satapathy AK, Pandey S, Chaudhary MR, Bagga A, Kabra M, Uwe K, Gupta N (2019) Report of another mutation proven case of carbonic anhydrase II deficiency. J Pediatr Genet 08:091–094
Seifter JL, Chang HY (2017) Disorders of acid-base balance: new perspectives . Kidney Dis (Basel) 2:170–186. https://doi.org/10.1159/000453028
Shah GN, Bonapace G, Hu PY, Strisciuglio P, Sly WS (2004a) Carbonic anhydrase II deficiency syndrome (osteopetrosis with renal tubular acidosis and brain calcification): Novel mutations in CA2 identified by direct sequencing expand the opportunity for genotype-phenotype correlation. Hum Mutat 24:272–272
Shah GN, Bonapace G, Hu PY, Strisciuglio P, Sly WS (2004b) Carbonic anhydrase II deficiency syndrome (osteopetrosis with renal tubular acidosis and brain calcification): novel mutations in CA2 identified by direct sequencing expand the opportunity for genotype-phenotype correlation. Hum Mutat 24:272. https://doi.org/10.1002/humu.9266
Silverman DN, McKenna R (2007) Solvent-mediated proton transfer in catalysis by carbonic anhydrase . Acc Chem Res 40:669–675. https://doi.org/10.1021/ar7000588
Sly WS et al (1985) Carbonic anhydrase II deficiency in 12 families with the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification . N Engl J Med 313:139–145
Sly WS, Hewett-Emmett D, Whyte MP, Yu YS, Tashian RE (1983) Carbonic anhydrase II deficiency identified as the primary defect in the autosomal recessive syndrome of osteopetrosis with renal tubular acidosis and cerebral calcification. Proc Natl Acad Sci USA 80:2752–2756. https://doi.org/10.1073/pnas.80.9.2752
Spicer SS, Lewis SE, Tashian RE, Schulte BA (1989) Mice carrying a CAR-2 null allele lack carbonic anhydrase II immunohistochemically and show vascular calcification. Am J Pathol 134:947–954
Supuran CT (2016) Structure and function of carbonic anhydrases. Biochem J 473:2023–2032. https://doi.org/10.1042/bcj20160115
The UniProtConsortium (2017) UniProt: the universal protein knowledgebase. Nucleic Acids Res 45:D158-d169. https://doi.org/10.1093/nar/gkw1099
Venta PJ, Welty RJ, Johnson TM, Sly WS, Tashian RE (1991) Carbonic anhydrase II deficiency syndrome in a Belgian family is caused by a point mutation at an invariant histidine residue (107 His----Tyr): complete structure of the normal human CA II gene American. J Hum Genet 49:1082–1090
Witt H et al (2006) A degradation-sensitive anionic trypsinogen (PRSS2) variant protects against chronic pancreatitis. Nat Genet 38:668–673. https://doi.org/10.1038/ng1797
Whyte MP (1993) Carbonic anhydrase II deficiency. Clin Orthop Relat Res (294):52–63
This study was approved by the ethics committee of Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital (2017 − 175).
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Yang, Y., Tang, N., Zhu, Y. et al. A novel homozygous nonsense mutation in the CA2 gene (c.368G>A, p.W123X) linked to carbonic anhydrase II deficiency syndrome in a Chinese family. Metab Brain Dis 36, 589–599 (2021). https://doi.org/10.1007/s11011-021-00677-9
- Carbonic anhydrase II deficiency syndrome
- Whole exome sequencing
- Homozygous nonsense mutation
- Minigene model
- Human embryonic kidney 293T cells