A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital Hypofibrinogenemia
We found a novel heterozygous mutation in the fibrinogen Bβ chain (c.490G>A) of a 3-year-old girl with congenital hypofibrinogenemia. To clarify the complex genetic mechanism, we made a mini-gene including a FGB c.490G>A mutation region, transfected it into a Chinese Hamster Ovary (CHO...
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doaj-82bb4e414a604d59898f07e1f43725ef2020-11-24T21:18:05ZengMDPI AGInternational Journal of Molecular Sciences1422-00672017-11-011811247010.3390/ijms18112470ijms18112470A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital HypofibrinogenemiaChiaki Taira0Kazuyuki Matsuda1Shinpei Arai2Mitsutoshi Sugano3Takeshi Uehara4Nobuo Okumura5Department of Health and Medical Sciences, Graduate School of Medicine, Shinshu University, 3-1-1 Asahi, Matsumoto 390-8621, JapanDepartment of Laboratory Medicine, Shinshu University Hospital, Matsumoto 390-8621, JapanDepartment of Laboratory Medicine, Shinshu University Hospital, Matsumoto 390-8621, JapanDepartment of Laboratory Medicine, Shinshu University Hospital, Matsumoto 390-8621, JapanDepartment of Laboratory Medicine, Graduate School of Medicine, Shinshu University, 3-1-1 Asahi, Matsumoto 390-8621, JapanDepartment of Health and Medical Sciences, Graduate School of Medicine, Shinshu University, 3-1-1 Asahi, Matsumoto 390-8621, JapanWe found a novel heterozygous mutation in the fibrinogen Bβ chain (c.490G>A) of a 3-year-old girl with congenital hypofibrinogenemia. To clarify the complex genetic mechanism, we made a mini-gene including a FGB c.490G>A mutation region, transfected it into a Chinese Hamster Ovary (CHO) cell line, and analyzed reverse transcription (RT) products. The assembly process and secretion were examined using recombinant mutant fibrinogen. Direct sequencing demonstrated that the mutant RT product was 99 bp longer than the wild-type product, and an extra 99 bases were derived from intron 3. In recombinant expression, a mutant Bβ-chain was weakly detected in the transfected CHO cell line, and aberrant fibrinogen was secreted into culture media; however, an aberrant Bβ-chain was not detected in plasma. Since the aberrant Bβ-chain was catabolized faster in cells, the aberrant Bβ-chain in a small amount of secreted fibrinogen may catabolize in the bloodstream. FGB c.490G>A indicated the activation of a cryptic splice site causing the insertion of 99 bp in intron 3. This splicing abnormality led to the production of a Bβ-chain possessing 33 aberrant amino acids, including two Cys residues in the coiled-coil domain. Therefore, a splicing abnormality may cause impaired fibrinogen assembly and secretion.https://www.mdpi.com/1422-0067/18/11/2470hypofibrinogenemiaFGBsplicing abnormalitycryptic splice site |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chiaki Taira Kazuyuki Matsuda Shinpei Arai Mitsutoshi Sugano Takeshi Uehara Nobuo Okumura |
spellingShingle |
Chiaki Taira Kazuyuki Matsuda Shinpei Arai Mitsutoshi Sugano Takeshi Uehara Nobuo Okumura A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital Hypofibrinogenemia International Journal of Molecular Sciences hypofibrinogenemia FGB splicing abnormality cryptic splice site |
author_facet |
Chiaki Taira Kazuyuki Matsuda Shinpei Arai Mitsutoshi Sugano Takeshi Uehara Nobuo Okumura |
author_sort |
Chiaki Taira |
title |
A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital Hypofibrinogenemia |
title_short |
A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital Hypofibrinogenemia |
title_full |
A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital Hypofibrinogenemia |
title_fullStr |
A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital Hypofibrinogenemia |
title_full_unstemmed |
A Novel Mutation in the Fibrinogen Bβ Chain (c.490G>A; End of Exon 3) Causes a Splicing Abnormality and Ultimately Leads to Congenital Hypofibrinogenemia |
title_sort |
novel mutation in the fibrinogen bβ chain (c.490g>a; end of exon 3) causes a splicing abnormality and ultimately leads to congenital hypofibrinogenemia |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2017-11-01 |
description |
We found a novel heterozygous mutation in the fibrinogen Bβ chain (c.490G>A) of a 3-year-old girl with congenital hypofibrinogenemia. To clarify the complex genetic mechanism, we made a mini-gene including a FGB c.490G>A mutation region, transfected it into a Chinese Hamster Ovary (CHO) cell line, and analyzed reverse transcription (RT) products. The assembly process and secretion were examined using recombinant mutant fibrinogen. Direct sequencing demonstrated that the mutant RT product was 99 bp longer than the wild-type product, and an extra 99 bases were derived from intron 3. In recombinant expression, a mutant Bβ-chain was weakly detected in the transfected CHO cell line, and aberrant fibrinogen was secreted into culture media; however, an aberrant Bβ-chain was not detected in plasma. Since the aberrant Bβ-chain was catabolized faster in cells, the aberrant Bβ-chain in a small amount of secreted fibrinogen may catabolize in the bloodstream. FGB c.490G>A indicated the activation of a cryptic splice site causing the insertion of 99 bp in intron 3. This splicing abnormality led to the production of a Bβ-chain possessing 33 aberrant amino acids, including two Cys residues in the coiled-coil domain. Therefore, a splicing abnormality may cause impaired fibrinogen assembly and secretion. |
topic |
hypofibrinogenemia FGB splicing abnormality cryptic splice site |
url |
https://www.mdpi.com/1422-0067/18/11/2470 |
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