Identification and functional characterization of novel polymorphisms associated with the genes for arylamine N-acetyltransferases in mice
Identification and functional characterization of novel polymorphisms associated with the genes for arylamine N-acetyltransferases in mice
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DOI：
10.1097/00008571-200207000-00006
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发表时间：
2002-07-01
期刊：
影响因子：
--
通讯作者：
Sim, E
中科院分区：
文献类型：
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作者：
Boukouvala, S;Price, N;Sim, E
人类芳胺N-乙酰转移酶（NAT）多态性与药物毒性和癌症易感性的变化有关。在小鼠中，三种NAT同工酶由Nat1、Nat2和Nat3基因编码。只有Nat 2以前已被证明是多态性的，单核苷酸取代导致A/J菌株中的慢乙酰化表型。我们对近交系（CBA和129/奥拉）、远交系（PO和TO）和野生近交系（Mus spretus和Mus musculus castaneus）小鼠的Nat基因进行了测序，并报告了M. spretus和M. M.栗色的肝匀浆酶活性测定表明，M。M. castaneus是一个'fast'，M. spretus是一种"缓慢"的乙酰化剂。Western blot分析表明，M. spretus比M. M.栗色的在哺乳动物细胞系中表达新的等位酶，并用一系列底物测量NAT酶活性。NAT1和NAT2同工酶。M.与M. castaneus相比，M. spretus。NAT3等位酶的活性几乎检测不到，尽管NAT3基因似乎确实被转录，因为在脾脏中通过RT-PCR检测到mRNA。已在BALB/c、C57 Bl/6 J、A/J、129/奥拉、CBA、PO、TO、M之间鉴定出对Nat相关遗传研究有用的其他多态性。M. castaneus和M. spretus菌株中位于Nat基因附近的4个微卫星重复序列。
Arylamine N-acetyltransferase (NAT) polymorphism in humans has been associated with variation in susceptibility to drug toxicity and cancer. In mice, three NAT isoenzymes are encoded by Nat1, Nat2 and Nat3 genes. Only Nat2 has been shown previously to be polymorphic, a single nucleotide substitution causing the slow acetylator phenotype in the A/J strain. We sequenced the Nat genes from inbred (CBA and 129/Ola), outbred (PO and TO) and wild-derived inbred (Mus spretus and Mus musculus castaneus) mouse strains and report polymorphism in all three Nat genes of M. spretus and in Nat2 and Nat3 genes of M. m. castaneus. Enzymatic activity assays using liver homogenates demonstrated that M. m. castaneus is a 'fast' and M. spretus a 'slow' acetylator. Western blot analysis indicated that hepatic NAT2 protein is less abundant in M. spretus than M. m. castaneus. The new allozymes were expressed in a mammalian cell line and NAT enzymatic activity was measured with a series of substrates. NAT1 and NAT2 isoenzymes of M. m. castaneus exhibited a higher rate of acetylation, compared with those of M. spretus. Activity of the NAT3 allozymes was hardly detectable, although the Nat3 gene does appear to be transcribed, since mRNA was detected by RT-PCR in the spleen. Additional polymorphisms, useful for Nat-related genetic studies, have been identified between BALB/c, C57Bl/6J, A/J, 129/ Ola, CBA, PO, TO, M. m. castaneus and M. spretus strains in four microsatellite repeats located close to the Nat genes.