Medical and Molecular Genetics, School of Clinical and Experimental Medicine and Centre for Rare Diseases and Personalised Medicine, University of Birmingham, Birmingham B15 2TT, UK.
Am J Hum Genet. 2011 Apr 8;88(4):499-507. doi: 10.1016/j.ajhg.2011.03.012.
Warburg Micro syndrome and Martsolf syndrome are heterogenous autosomal-recessive developmental disorders characterized by brain, eye, and endocrine abnormalities. Previously, identification of mutations in RAB3GAP1 and RAB3GAP2 in both these syndromes implicated dysregulation of the RAB3 cycle (which controls calcium-mediated exocytosis of neurotransmitters and hormones) in disease pathogenesis. RAB3GAP1 and RAB3GAP2 encode the catalytic and noncatalytic subunits of the hetrodimeric enzyme RAB3GAP (RAB3GTPase-activating protein), a key regulator of the RAB3 cycle. We performed autozygosity mapping in five consanguineous families without RAB3GAP1/2 mutations and identified loss-of-function mutations in RAB18. A c.71T > A (p.Leu24Gln) founder mutation was identified in four Pakistani families, and a homozygous exon 2 deletion (predicted to result in a frameshift) was found in the fifth family. A single family whose members were compound heterozygotes for an anti-termination mutation of the stop codon c.619T > C (p.X207QextX20) and an inframe arginine deletion c.277_279 del (p.Arg93 del) were identified after direct gene sequencing and multiplex ligation-dependent probe amplification (MLPA) of a further 58 families. Nucleotide binding assays for RAB18(Leu24Gln) and RAB18(Arg93del) showed that these mutant proteins were functionally null in that they were unable to bind guanine. The clinical features of Warburg Micro syndrome patients with RAB3GAP1 or RAB3GAP2 mutations and RAB18 mutations are indistinguishable, although the role of RAB18 in trafficking is still emerging, and it has not been linked previously to the RAB3 pathway. Knockdown of rab18 in zebrafish suggests that it might have a conserved developmental role. Our findings imply that RAB18 has a critical role in human brain and eye development and neurodegeneration.
沃伯格微综合征和马索夫综合征是两种异质性常染色体隐性发育障碍,其特征为脑、眼和内分泌异常。先前,在这两种综合征中发现 RAB3GAP1 和 RAB3GAP2 突变,提示 RAB3 循环(控制神经递质和激素的钙介导的胞吐作用)失调在疾病发病机制中起作用。RAB3GAP1 和 RAB3GAP2 编码异二聚体酶 RAB3GAP(RAB3GTPase 激活蛋白)的催化和非催化亚基,是 RAB3 循环的关键调节剂。我们在 5 个无 RAB3GAP1/2 突变的近亲家庭中进行了纯合子作图,并在 RAB18 中发现了功能丧失突变。在 4 个巴基斯坦家庭中发现了 c.71T > A(p.Leu24Gln)的起始突变,在第 5 个家庭中发现了纯合子外显子 2 缺失(预计会导致移码)。在直接基因测序和进一步的 58 个家庭的多重连接依赖性探针扩增(MLPA)后,发现了一个家族的成员为终止密码子 c.619T > C(p.X207QextX20)的抗终止突变和内含子 277_279del(p.Arg93del)的复合杂合子。RAB18(Leu24Gln)和 RAB18(Arg93del)的核苷酸结合测定表明,这些突变蛋白在功能上是无效的,因为它们无法结合鸟嘌呤。尽管 RAB18 在运输中的作用仍在出现,并且以前尚未与 RAB3 途径相关联,但具有 RAB3GAP1 或 RAB3GAP2 突变和 RAB18 突变的沃伯格微综合征患者的临床特征无法区分。斑马鱼中 rab18 的敲低表明它可能具有保守的发育作用。我们的发现表明,RAB18 在人类大脑和眼睛发育及神经退行性变中具有关键作用。