IPATIMUP - Institute of Pathology and Molecular Immunology of University of Porto, Rua Dr Roberto Frias s/n, 4200-465, Porto, Portugal.
J Inherit Metab Dis. 2008 Dec;31 Suppl 2:S457-60. doi: 10.1007/s10545-008-1046-z. Epub 2008 Dec 16.
Maple syrup urine disease (MSUD) is a rare disorder of branched-chain amino acid (BCAA) metabolism caused by the defective function of branched-chain α-ketoacid dehydrogenase complex (BCKD). Many MSUD-causing mutations have already been described in genes that encode the complex (BCKDHA, BCKDHB and DBT), but up to now only four large deletions are known, all located in the DBT gene. In a previous study we identified a Portuguese MSUD patient with a homozygous deletion of exons 2, 3 and 4 at the BCKDHA gene; however, the corresponding breakpoints and, consequently, the exact deletion extension were not identified. Here, using long-range PCR and sequencing methodologies we were able to refine the characterization of this gross rearrangement. A genomic DNA loss of about 13.8 kb was detected, starting at intron 1 and ending at intron 4, thus encompassing exons 2, 3 and 4. Molecular characterization showed that the deletion junction contained a short sequence whose motif was CGGG. Since this motif is present in introns 1 and 4 of normal genomic DNA, we have hypothesized that non-homologous recombination was the mechanism underlying the identified large deletion, within which the CGGG could be derived either from intron 1 or from intron 4.
枫糖尿症(MSUD)是一种罕见的支链氨基酸(BCAA)代谢紊乱,由支链α-酮酸脱氢酶复合物(BCKD)的功能缺陷引起。许多导致 MSUD 的突变已经在编码该复合物的基因(BCKDHA、BCKDHB 和 DBT)中被描述,但到目前为止,只知道四种大的缺失,都位于 DBT 基因中。在之前的一项研究中,我们鉴定了一名葡萄牙 MSUD 患者,其 BCKDHA 基因的外显子 2、3 和 4 发生纯合缺失;然而,相应的断点以及确切的缺失扩展没有被确定。在这里,我们使用长距离 PCR 和测序方法能够对这种严重的重排进行精细化特征描述。从内含子 1 到内含子 4 检测到大约 13.8 kb 的基因组 DNA 缺失,因此包含外显子 2、3 和 4。分子特征显示,缺失连接点包含一段短序列,其基序为 CGGG。由于该基序存在于正常基因组 DNA 的内含子 1 和 4 中,我们假设非同源重组是导致所鉴定的大片段缺失的机制,其中的 CGGG 可能来自内含子 1 或内含子 4。