McDowell M L, Nguyen T, Cleaver J E
Laboratory of Radiobiology and Environmental Health, University of California, San Francisco 94143-0750.
Mutagenesis. 1993 Mar;8(2):155-61. doi: 10.1093/mutage/8.2.155.
The XPAC (xeroderma pigmentosum group A complementing) gene, which is located on chromosome 9, carries a variety of point mutations in XP group A patients. We investigated the role of the XPAC gene product in excision repair by generating revertants of an XP group A cell line (XP12RO) that have increased resistance to ultraviolet light. One of these cell lines, XP129, can repair (6-4) pyrimidine-pyrimidone photoproducts normally but has reduced repair of cyclobutane dimers, as in XP12RO. Sequence analysis of cDNA from the XPAC gene indicated that XP12RO contains a termination codon at amino acid position 207, resulting in a reduced amount of mRNA and no detectable protein. In the revertant XP129 line, this termination codon has been mutated further and now encodes glycine in one allele instead of the wild-type arginine. The mRNA level detected by allele-specific polymerase chain reaction amplification was greater for the reverted sequence than for the chain-terminating sequence. These observations indicated that a point mutation resulting in a mis-sense mutation in the XPAC gene and altered expression of the XPAC protein can alter the substrate specificity of the excision repair system, and imply that the XPAC gene product plays an important role in photoproduct recognition.
着色性干皮病 A 互补(XPAC)基因位于 9 号染色体上,在 A 型着色性干皮病患者中存在多种点突变。我们通过构建对紫外线具有更高抗性的 A 型着色性干皮病细胞系(XP12RO)的回复突变体,研究了 XPAC 基因产物在切除修复中的作用。其中一个细胞系 XP129,能正常修复(6 - 4)嘧啶 - 嘧啶酮光产物,但对环丁烷二聚体的修复能力降低,与 XP12RO 情况相同。对 XPAC 基因的 cDNA 进行序列分析表明,XP12RO 在氨基酸位置 207 处含有一个终止密码子,导致 mRNA 量减少且未检测到蛋白质。在回复突变体 XP129 细胞系中,这个终止密码子进一步发生突变,现在一个等位基因编码甘氨酸而非野生型精氨酸。通过等位基因特异性聚合酶链反应扩增检测到的回复序列的 mRNA 水平高于链终止序列。这些观察结果表明,XPAC 基因中的一个点突变导致错义突变并改变了 XPAC 蛋白的表达,可改变切除修复系统的底物特异性,这意味着 XPAC 基因产物在光产物识别中起重要作用。