A functionally critical single nucleotide polymorphism in the gene encoding the membrane-bound alcohol dehydrogenase found in ethanol oxidation-deficient Gluconobacter thailandicus
A functionally critical single nucleotide polymorphism in the gene encoding the membrane-bound alcohol dehydrogenase found in ethanol oxidation-deficient Gluconobacter thailandicus
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在乙醇氧化缺陷的泰国葡糖杆菌中发现编码膜结合乙醇脱氢酶的基因中的功能关键单核苷酸多态性
DOI：
10.1016/j.gene.2015.04.080
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发表时间：
2015
期刊：
影响因子：
3.5
通讯作者：
K Matsushita
中科院分区：
文献类型：
--
作者：
P Charoenyingcharoen;M Matsutani;T Yakushi;G Theeragool;P Yukphan;K Matsushita
泰国葡萄糖酸杆菌菌株NBRC 3254、NBRC 3255、NBRC 3256、NBRC 3257和NBRC 3258在乙醇氧化呼吸链中是天然缺陷的，因为它们不产生膜结合乙醇脱氢酶（ADH）的细胞色素亚基。G.泰国菌株NBRC 3255和NBRC 3257与公共数据库中的一个密切相关的基因相比，编码细胞色素亚基的adhB基因存在4个碱基差异，其中一个核苷酸差异导致信号序列中第-19位色氨酸（Trp）的Opal密码子转移到周质空间。（此处，+1残基的位置被指定为信号肽切割后的N-末端氨基酸残基），而其它差异导致一个错义和两个沉默氨基酸改变。所有五个G。Thailandicus菌株具有Trp（-19）Opal突变。乙醇氧化和ADH活性在NBRC 3255中恢复与内源adhB基因的衍生物，其中-19欧泊密码子被改变编码色氨酸的转化。这些结果表明，该序列是一个功能关键的单核苷酸多态性的细胞色素亚基。对NBRC 3255和NBRC 3257的基因组草图进行的比较基因组分析显示，尽管这两个基因组密切相关，但它们都有大量独特的开放阅读框架。我们认为，密切相关的NBRC 3255和NBRC 3257从一个共同的祖先有突变的theadhB基因，而没有额外的功能关键突变发生在theadhB假基因在进化过程中。
The Gluconobacterthailandicusstrains NBRC3254, NBRC3255, NBRC3256, NBRC3257, and NBRC3258 are naturally deficient in the ethanol-oxidizing respiratory chain because they do not produce the cytochrome subunit of the membrane-bound alcohol dehydrogenase (ADH). Draft genomes ofG. thailandicusstrains NBRC3255 and NBRC3257 indicated that theadhBgene encoding the cytochrome subunit contains four base differences when compared to a closely related gene in the public database One of the nucleotide differences results in an Opal codon at the –19th tryptophan (Trp) in the signal sequence for translocation to the periplasmic space (here, the position of + 1st residue is assigned to the N-terminal amino acid residue after signal peptide cleavage), while the other differences result in one missense and two silent amino acid alterations. All five of theG. thailandicusstrains were shown to have the Trp(-19)Opal alteration. Ethanol oxidation and ADH activities in NBRC3255 were restored by transformation with a derivative of the endogenousadhBgene, of which the –19th Opal codon was altered to encode Trp. These results indicate that this sequence is a functionally critical single nucleotide polymorphism in the cytochrome subunit. Comparative genomic analyses between the draft genomes of NBRC3255 and NBRC3257 revealed that although the two genomes are closely related, they both have a significant number of unique open reading frames. We suggest that the closely related NBRC3255 and NBRC3257 diverged from a common ancestor having the mutation in theadhBgene, whereas no additional functionally critical mutation occurred in theadhBpseudogene over the course of evolution.