Matejas Verena, Huehne Kathrin, Thiel Christian, Sommer Claudia, Jakubiczka Sibylle, Rautenstrauss Bernd
Institute of Human Genetics, Friedrich Alexander University, Erlangen, Germany.
Neurogenetics. 2006 May;7(2):119-26. doi: 10.1007/s10048-006-0030-8. Epub 2006 Mar 29.
Hereditary neuropathy with liability to pressure palsies (HNPP) is most frequently caused by deletion of a 1.4-Mb region in chromosome 17p11.2-12 including the peripheral myelin protein 22 (PMP22) gene. Smaller deletions partially affecting the PMP22 gene are less frequently observed. We identified in a HNPP patient a deletion of the 5' region of PMP22 including non-coding exon 1, coding exons 2 and 3, whereas, exons 4 and 5 were present. PMP22 exon 3- and 4-specific qPCR resulted in a deletion of one exon 3 allele but in the presence of 2 exon 4 alleles. SNP analysis revealed the presence of heterozygosity for PMP22 coding exons 4 and 5. Finally, MLPA specific for the CMT1A region defined this deletion for the entire 5' region of PMP22 (exons 1, 2 and 3). These partial HNPP deletions may be missed by other techniques, e.g., STR marker analysis. Alu elements have been reported to mediate non-allelic recombination events. Bioinformatic analysis revealed 12 Alu elements flanking in close neighbourhood the estimated 40-kb deletion region as candidates for recombination events. PCR primers were designed to identify a breakpoint-spanning product including the respective Alu elements. PCR-driven identification of a junction fragment was successful with AluJo-AluSq and AluYb9-AluSq specific primer pairs comprising the same intronic region of PMP22. Sequence analysis of these breakpoint-overlapping PCR fragments revealed a 29-bp motif including a chi-like sequence (GCTGG) present both in the AluYb9 and the AluSq element. These data confirm that low-copy repeats (LCRs) mediate non-allelic homologous recombinations (NAHR).
遗传性压力易感性周围神经病(HNPP)最常见的病因是17号染色体p11.2 - 12区域一个1.4 Mb片段的缺失,该区域包含外周髓鞘蛋白22(PMP22)基因。较少观察到部分影响PMP22基因的较小缺失。我们在一名HNPP患者中鉴定出PMP22基因5'区域的缺失,包括非编码外显子1、编码外显子2和3,而外显子4和5存在。PMP22外显子3和4特异性定量PCR结果显示一个外显子3等位基因缺失,但存在2个外显子4等位基因。单核苷酸多态性(SNP)分析显示PMP22编码外显子4和5存在杂合性。最后,针对遗传性运动感觉神经病1型A(CMT1A)区域的多重连接探针扩增(MLPA)确定了PMP22整个5'区域(外显子1、2和3)的这种缺失。其他技术,如短串联重复序列(STR)标记分析,可能会遗漏这些部分HNPP缺失。据报道,Alu元件介导非等位基因重组事件。生物信息学分析显示,在估计的40 kb缺失区域附近紧密侧翼排列着12个Alu元件,作为重组事件的候选元件。设计了聚合酶链反应(PCR)引物来鉴定包含相应Alu元件的跨越断点的产物。使用包含PMP22相同内含子区域的AluJo - AluSq和AluYb9 - AluSq特异性引物对,通过PCR成功鉴定出一个连接片段。对这些跨越断点的PCR片段进行序列分析,发现一个29 bp的基序,其中包含一个在AluYb9和AluSq元件中都存在的类chi序列(GCTGG)。这些数据证实低拷贝重复序列(LCRs)介导非等位基因同源重组(NAHR)。