Propionic acidemia : Molecular analysis of beta subnit deficient Japanese petients

丙酸血症：β亚基缺陷日本患者的分子分析

• 批准号: 04670575
• 负责人: OHURA Toshihiro
• 金额: $ 1.22万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670575/
• 关键词: Propionic Acidemia; Propionyl CoA Ccrboxylase; Exon skipping

Propionic acidemia is a recessively inherited disorder of organic acid metabolism caused by deficiency of propionyl-CoA carboxylase (PCC) activity. Enxyme deficiency can result from mutations in either the alpha or the beta subunit. The human beta-PCC cDNA was sequenced in full and shown to encode a pre-beta subunit of 539 amino acids. Polymerase chain reaction amplification and sequencing of betaPCC cDNA from five beta-subunit deficient Japanese patients (cell no.83,187,212,276,338) revealed two missense mutations (C1283T, C493T), one nonsense mutation (C1495T) and two splicing mutations. The allele frequency of the C1283T mutation was 50% (5/10) in this study, and this mutation was not detected in 50 normal Japanese subjects. This suggests that the C1283T mutation causes disease, and may be a common mutation in Japanese propionic acidemia patients. Two deletions were found in the coding region of the beta-PCC cDNA.One is a 57-bp in-frame deletion between nucleothides 373 and 429, resulting in the deletion of 19 amino acids in the coding sequence in patient 187. Analysis of the genomic DNA from 187 revealed a 4-bp deletion from bp+3 to bp+6 of the downstream intron adjacent to the deleted exon. This deletion disrupted the consensus 5' splice signal (GTAAGT->GTGTTT) and led to exon skipping. The second deletion is a 101-bp deletion between nucleotides 1199 and 1299 in patient 338, which resulted in a frame shift and a stop codon in the new frame. Similar analysis of the genomic DNA from 338 revealed an 8-bp deletion from bp+3 to bp+10 of the donor splice site of the intron. This deletion also disrupted the consensus 5' splice signal (GTGAGG->GTCATG) and led to exon skipping.

丙酸血症是由丙酰辅酶A羧化酶（PCC）活性缺乏引起的一种遗传性有机酸代谢紊乱。酶缺乏可由α或β亚基的突变引起。对人β-PCC cDNA进行了全序列测定，并显示其编码539个氨基酸的前β亚基。对5例日本β-亚单位缺陷患者（细胞编号83、187、212、276、338）的β-PCC cDNA进行聚合酶链反应扩增和测序，发现2个错义突变（C1283 T、C493 T）、1个无义突变（C1495 T）和2个剪接突变。本研究中C1283 T突变的等位基因频率为50%（5/10），在50名日本正常受试者中未检测到该突变。这表明C1283 T突变引起疾病，并且可能是日本丙酸血症患者中常见的突变。在β-PCC cDNA编码区发现两处缺失，一处是位于第373和429号核苷酸之间的57-bp框内缺失，导致187号患者编码序列中19个氨基酸的缺失。对187个基因组DNA的分析显示，与缺失的外显子相邻的下游内含子从bp+3到bp+6有4个碱基的缺失。这种缺失破坏了共有5'剪接信号（GTAAGT->GTGTTT）并导致外显子跳跃。第二个缺失是患者338中核苷酸1199和1299之间的101-bp缺失，这导致新框架中的移码和终止密码子。对338的基因组DNA进行类似的分析，发现内含子的供体剪接位点从bp+3到bp+10有一个8-bp的缺失。这种缺失也破坏了共有5'剪接信号（GTGAGG->GTCATG）并导致外显子跳跃。

项目成果

OHURA,T.et al: "Propionic acidemia:Sequence analysis of mutant mRNAs from Japanese β subunit-deficient patients." Journal of Inherited Metabolic Disease. 16. (1993)

OHURA, T. 等人：“丙酸血症：来自日本 β 亚基缺陷患者的突变 mRNA 的序列分析。遗传代谢疾病杂志 16。”

Ohura J.,et al.: "The molecular defect in propionic acidemia:exon skipping eaused by on 8-bp deletion from on citron in the PCCB allele." Hum.Genet.92. 397-402 (1993)

Ohura J.,et al.：“丙酸血症的分子缺陷：PCCB 等位基因中柠檬子上的 8 bp 缺失导致外显子跳跃。”

大浦 敏博: "プロピオン酸血症" 臨床検査. 36. 233 (1992)

Toshihiro Oura：“丙酸血症”临床检查。36。233（1992）。

Ohura T.et al: "The molecular defect in propionic acidemia : Exon skipping caused by on 8-bp deletion from an intron in the PCC B allele" Hum. Genet.Vol.92. 397-402 (1993)

Ohura T.等人：“丙酸血症的分子缺陷：PCC B 等位基因内含子的 8 bp 缺失导致外显子跳跃”Hum。

Jahara J.,et al.: "Three independent mutation in the same exon of the PCCB gene:Differences between Cancasians and Japanese propionic acidemia." Journal of Inherited Metabolic Disease. 16. 353-360 (1993)

Jahara J.,et al.：“PCCB 基因同一外显子中的三个独立突变：加拿大人与日本丙酸血症之间的差异。”