Tager J M, Brul S, Wiemer E A, Strijland A, Van Driel R, Schutgens R B, Van den Bosch H, Wanders R J, Westerveld A
Laboratory of Biochemistry, University of Amsterdam, The Netherlands.
Prog Clin Biol Res. 1990;321:545-58.
The peroxisomal diseases can be divided into three categories: 1) diseases in which morphologically distinguishable peroxisomes are virtually absent (Zellweger syndrome; infantile Refsum disease; Hyperpipecolic Acidaemia; neonatal Adrenoleukodystrophy); 2) diseases in which peroxisomes are present but several peroxisomal functions are impaired (rhizomelic Chondrodysplasia punctata; Zellweger-like syndrome?); and 3) diseases in which a single peroxisomal function is impaired. We have used complementation analysis after somatic cell fusion in order to investigate the genetic relationship between diseases in category 1. The activity of acyl-CoA: dihydroxyacetonephosphate acyltransferase, which is deficient in these diseases and in rhizomelic Chondrodysplasia punctata, was used as an index of complementation. The cell lines studied, all of which complemented with rhizomelic Chondrodysplasia punctata, could be divided into at least 4 and possibly 5 complementation groups. This indicates that at least 5 and possibly 6 genes are involved in the assembly of peroxisomes. One of the complementation groups is comprised of cell lines from patients with the Zellweger syndrome, infantile Refsum disease and Hyperpipecolic Acidaemia. Thus mutations in the same gene can lead to clinically distinguishable diseases. On the other hand, the Zellweger cell lines studied fall into 3 complementation groups and the infantile Refsum disease cell lines into 2 groups. Thus mutations in different genes can lead to the same clinical phenotype. Fusion of complementary cell lines lacking morphologically distinguishable peroxisomes leads to assembly of peroxisomes, which can be monitored by measuring particle-bound catalase biochemically or by immunofluorescence. In two combinations of cell lines assembly of peroxisomes was rapid and cycloheximide insensitive. Thus the components required for peroxisome assembly must be present in a stable form in the parental cell lines, at least one of which must contain peroxisomal ghost-like structures.
1）几乎不存在形态上可区分的过氧化物酶体的疾病（泽尔韦格综合征；婴儿型雷夫叙姆病；高哌可酸血症；新生儿肾上腺脑白质营养不良）；2）存在过氧化物酶体但几种过氧化物酶体功能受损的疾病（肢根型点状软骨发育不良；类泽尔韦格综合征？）；以及3）单一过氧化物酶体功能受损的疾病。我们利用体细胞融合后的互补分析来研究第1类疾病之间的遗传关系。在这些疾病和肢根型点状软骨发育不良中缺乏的酰基辅酶A：磷酸二羟丙酮酰基转移酶的活性被用作互补的指标。所研究的细胞系，所有这些细胞系都能与肢根型点状软骨发育不良互补，可分为至少4个，可能5个互补组。这表明至少5个，可能6个基因参与过氧化物酶体的组装。其中一个互补组由来自泽尔韦格综合征、婴儿型雷夫叙姆病和高哌可酸血症患者的细胞系组成。因此，同一基因的突变可导致临床上可区分的疾病。另一方面，所研究的泽尔韦格细胞系分为3个互补组，婴儿型雷夫叙姆病细胞系分为2个组。因此，不同基因的突变可导致相同的临床表型。缺乏形态上可区分的过氧化物酶体的互补细胞系融合会导致过氧化物酶体的组装，这可以通过生化测量颗粒结合过氧化氢酶或通过免疫荧光来监测。在两种细胞系组合中，过氧化物酶体的组装迅速且对放线菌酮不敏感。因此，过氧化物酶体组装所需的成分必须以稳定的形式存在于亲代细胞系中，其中至少一个细胞系必须含有过氧化物酶体样的结构。
