A Novel Intronic Splicing Mutation in the EXT2 Gene of a Chinese Family with Multiple Osteochondroma

被引:3
作者
Guo, Xiaoyan [1 ]
Chen, Shunyou [2 ]
Lin, Mingrui [4 ]
Pan, Yuancheng [2 ]
Liu, Nannan [3 ]
Shi, Tengfei [1 ]
机构
[1] Xiamen Univ, Dept Lab Med, Fuzhou Hosp Affiliated 2, 47 Shangteng Rd, Fuzhou 350007, Fujian, Peoples R China
[2] Xiamen Univ, Dept Orthoped, Fuzhou Hosp Affiliated 2, Fuzhou, Peoples R China
[3] Xiamen Univ, Orthoped Inst, Fuzhou Hosp Affiliated 2, Fuzhou, Peoples R China
[4] Fujian Tradit Med Univ, Intens Care Unit, Affiliated Peoples Hosp, Fuzhou, Peoples R China
关键词
multiple osteochondroma; EXT2; gene; splicing mutation; bioinformatics; hybrid minigene splicing assay; GENOTYPE-PHENOTYPE CORRELATION; HEPARAN-SULFATE; NATURAL-HISTORY; IDENTIFICATION; GROWTH; HETEROZYGOSITY; CHONDROGENESIS; MECHANISMS; PROTEINS; SITE;
D O I
10.1089/gtmb.2021.0030
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Background: Multiple osteochondroma (MO), an autosomal dominant genetic disease, is caused by heterozygous mutations in the EXT1 and EXT2 genes. Approximately 80% of pathogenic mutations are nonsense/missense mutations, small indels, and splicing mutations. Splicing mutations, particularly at the 3 ' and 5 ' splice sites, disrupt normal mRNA processing and cause exon skipping or aberrant splicing, ultimately resulting in protein truncation and loss of function. Methods: Polymerase chain reaction (PCR) and Sanger sequencing were applied to detect subtle mutations in a Chinese family with MO, the pathogenicity of a splicing variant was predicted by bioinformatics and further verified using a minigene splicing assay. Results: A novel and heterozygous splicing mutation, c.626 + 2_626 + 5delTAGG, was identified in the EXT2 gene of the proband and the father by PCR and Sanger sequencing, whereas the unaffected mother and brother had wild-type alleles at the same site. Bioinformatics predicted that the 5 ' splicing site of exon 3 in the EXT2 gene was destroyed due to this mutation. A hybrid minigene splicing assay (HMSA) indicated that the mutation disturbed the normal splicing of the EXT2 gene mRNA and led to a deletion of 79 bp at the 5 ' end of exon 3, which resulted in aberrant splicing of exon 3 and introduced an earlier stop codon in the EXT2 gene. Conclusion: A novel splicing mutation was identified that produced the MO phenotype through aberrant splicing in a Chinese family. This observation, expands our knowledge of the spectrum of molecular pathogenic mechanisms leading to aberrant mRNA splicing.
引用
收藏
页码:478 / 485
页数:8
相关论文
共 38 条
  • [1] Splicing mutations in human genetic disorders: examples, detection, and confirmation
    Abramowicz, Anna
    Gos, Monika
    [J]. JOURNAL OF APPLIED GENETICS, 2018, 59 (03) : 253 - 268
  • [2] CLONING OF THE PUTATIVE TUMOR-SUPPRESSOR GENE FOR HEREDITARY MULTIPLE EXOSTOSES (EXT1)
    AHN, J
    JOSEFLUDECKE, H
    LINDOW, S
    HORTON, WA
    LEE, B
    WAGNER, MJ
    HORSTHEMKE, B
    WELLS, DE
    [J]. NATURE GENETICS, 1995, 11 (02) : 137 - 143
  • [3] Identification of a mutation that perturbs NF1 gene splicing using genomic DNA samples and a minigene assay
    Baralle, M
    Baralle, D
    De Conti, L
    Mattocks, C
    Whittaker, J
    Knezevich, A
    Ffrench-Constant, C
    Baralle, FE
    [J]. JOURNAL OF MEDICAL GENETICS, 2003, 40 (03) : 220 - 222
  • [4] Fatigue and pain in children and adults with multiple osteochondromas in Norway, a cross-sectional study
    Bathen, Trine
    Fredwall, Svein
    Steen, Unni
    Svendby, Ellen Berg
    [J]. INTERNATIONAL JOURNAL OF ORTHOPAEDIC AND TRAUMA NURSING, 2019, 34 : 28 - 35
  • [5] Interactions of signaling proteins, growth factors and other proteins with heparan sulfate: mechanisms and mysteries
    Billings, Paul C.
    Pacifici, Maurizio
    [J]. CONNECTIVE TISSUE RESEARCH, 2015, 56 (04) : 272 - 280
  • [6] Mechanisms of alternative pre-messenger RNA splicing
    Black, DL
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 2003, 72 : 291 - 336
  • [7] EXT-mutation analysis and loss of heterozygosity in sporadic and hereditary osteochondromas and secondary chondrosarcomas
    Bovée, JVMG
    Cleton-Jansen, AM
    Wuyts, W
    Caethoven, G
    Taminiau, AHM
    Bakker, E
    Van Hul, W
    Cornelisse, CJ
    Hogendoorn, PCW
    [J]. AMERICAN JOURNAL OF HUMAN GENETICS, 1999, 65 (03) : 689 - 698
  • [8] The extostosin family: Proteins with many functions
    Busse-Wicher, Marta
    Wicher, Krzysztof B.
    Kusche-Gullberg, Marion
    [J]. MATRIX BIOLOGY, 2014, 35 : 25 - 33
  • [9] Forloni M, 2018, COLD SPRING HARB PRO, V1, P663
  • [10] Genotype-phenotype correlation in hereditary multiple exostoses
    Francannet, C
    Cohen-Tanugi, A
    Le Merrer, M
    Munnich, A
    Bonaventure, J
    Legeai-Mallet, L
    [J]. JOURNAL OF MEDICAL GENETICS, 2001, 38 (07) : 430 - 434