Hayashi S, Watanabe J, Nakachi K, Kawajiri K
Department of Biochemistry, Saitama Cancer Center Research Institute.
J Biochem. 1991 Sep;110(3):407-11. doi: 10.1093/oxfordjournals.jbchem.a123594.
Individuals with high genetic risk of lung cancer had previously been identified by MspI polymorphisms of the cytochrome P450IA1 gene. In the present study we analyzed the structures of individual P450IA1 genes by PCR direct sequencing of genomic DNA of each genotype raised by the MspI polymorphisms, which were ascribed to a single point mutation in the 3'-flanking region. We then found a novel point mutation in the coding region of the gene which results in the substitution of Ile for Val at residue 462 in the heme binding region. We further analyzed the genetic association between this amino acid replacement and MspI polymorphisms in the general population, using a new method to detect polymorphisms not recognized by restriction enzymes. The results showed that there are at least two forms of human P450IA1 protein with different primary structures and that one of the forms is closely linked with the lung cancer-susceptible genotype of MspI polymorphisms. Thus MspI polymorphisms, which are associated with increased risk of lung cancer, are linked to at least one amino acid substitution, which gives an important clue, at the molecular level, toward elucidation of increased susceptibility to lung cancer.
肺癌高遗传风险个体此前已通过细胞色素P450IA1基因的MspI多态性得以鉴定。在本研究中,我们通过对由MspI多态性产生的每种基因型的基因组DNA进行PCR直接测序,分析了个体P450IA1基因的结构,这些多态性归因于3'侧翼区域的单个点突变。然后我们在该基因的编码区发现了一个新的点突变,该突变导致血红素结合区域第462位残基处的异亮氨酸取代缬氨酸。我们使用一种检测限制酶无法识别的多态性的新方法,进一步分析了普通人群中这种氨基酸替换与MspI多态性之间的遗传关联。结果表明,人类P450IA1蛋白至少有两种具有不同一级结构的形式,其中一种形式与MspI多态性的肺癌易感基因型密切相关。因此,与肺癌风险增加相关的MspI多态性与至少一种氨基酸替换有关,这在分子水平上为阐明肺癌易感性增加提供了重要线索。