Genetic and metabolomic analysis of AdeD and AdeI mutants of de novo purine biosynthesis: cellular models of de novo purine biosynthesis deficiency disorders.
Genetic and metabolomic analysis of AdeD and AdeI mutants of de novo purine biosynthesis: cellular models of de novo purine biosynthesis deficiency disorders.
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DOI：
10.1016/j.ymgme.2013.01.002
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发表时间：
2013-03
影响因子：
3.8
通讯作者：
Patterson, David
中科院分区：
文献类型：
--
作者：
Duval, Nathan;Luhrs, Kyleen;Wilkinson, Terry G., II;Baresova, Veronika;Skopova, Vaclava;Kmoch, Stanislav;Vacano, Guido N.;Zikanova, Marie;Patterson, David
关键词：
嘌呤是许多细胞过程所必需的分子，包括RNA和DNA合成、酶活性调节、蛋白质合成和功能、能量代谢和转移、必需辅酶功能和细胞信号传导。嘌呤通过嘌呤从头生物合成途径产生。嘌呤生物合成基因，例如磷酸核糖氨基咪唑羧化酶/磷酸核糖氨基咪唑琥珀酰胺合成酶（PAICS，E.C. 6.3.2.6/E.C. 4.1.1.21），可导致低等脊椎动物的发育异常。人类PAICS表达的改变与各种类型的癌症有关。腺苷酸琥珀酸裂解酶（ADSL，E.C. 4.3.2.2）或5-氨基咪唑-4-甲酰胺核糖核苷酸甲酰基转移酶/IMP环化水解酶（ATIC，E.C. 2.1.2.3/E.C. 3.5.4.10）导致具有一系列临床症状的先天性代谢缺陷，包括发育迟缓、严重的神经症状、肾结石、联合免疫缺陷和自闭症特征。这些疾病的发病机制尚不清楚，也没有有效的治疗方法。对携带各种从头嘌呤生物合成途径基因突变的细胞的研究提供了一种分析嘌呤疾病的方法。在这里，我们报告的AdeD中国仓鼠卵巢（CHO）细胞，携带基因突变编码p.E177K和p.W363* 变体的PAICS的表征。两种突变都影响PAICS结构并完全消除其生物合成。此外，我们描述了一种灵敏和快速的分析方法检测嘌呤从头生物合成中间体的基础上，高效液相色谱电化学检测。使用这种技术，我们检测到积累的空气在AdeD细胞。在AdeI细胞中，ADSL基因的突变体，我们检测到SAICAR和SAMP的积累，有点出乎意料的是，AIR的积累。该方法具有很大的潜力，从头嘌呤生物合成途径突变体的代谢物谱，在人类中的嘌呤代谢的新的遗传缺陷的鉴定，并阐明这一关键的代谢途径的调节。
Purines are molecules essential for many cell processes, including RNA and DNA synthesis, regulation of enzyme activity, protein synthesis and function, energy metabolism and transfer, essential coenzyme function, and cell signaling. Purines are produced via the de novo purine biosynthesis pathway. Mutations in purine biosynthetic genes, for example phosphoribosylaminoimidazole carboxylase/phosphoribosylaminoimidazole succinocarboxamide synthetase (PAICS, E.C. 6.3.2.6/E.C. 4.1.1.21), can lead to developmental anomalies in lower vertebrates. Alterations in PAICS expression in humans have been associated with various types of cancer. Mutations in adenylosuccinate lyase (ADSL, E.C. 4.3.2.2) or 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC, E.C. 2.1.2.3/E.C. 3.5.4.10) lead to inborn errors of metabolism with a range of clinical symptoms, including developmental delay, severe neurological symptoms, renal stones, combined immunodeficiency, and autistic features. The pathogenetic mechanism is unknown for any of these conditions, and no effective treatments exist. The study of cells carrying mutations in the various de novo purine biosynthesis pathway genes provides one approach to analysis of purine disorders. Here we report the characterization of AdeD Chinese hamster ovary (CHO) cells, which carry genetic mutations encoding p.E177K and p.W363* variants of PAICS. Both mutations impact PAICS structure and completely abolish its biosynthesis. Additionally, we describe a sensitive and rapid analytical method for detection of purine de novo biosynthesis intermediates based on high performance liquid chromatography with electrochemical detection. Using this technique we detected accumulation of AIR in AdeD cells. In AdeI cells, mutant for the ADSL gene, we detected accumulation of SAICAR and SAMP and, somewhat unexpectedly, accumulation of AIR. This method has great potential for metabolite profiling of de novo purine biosynthesis pathway mutants, identification of novel genetic defects of purine metabolism in humans, and elucidating the regulation of this critical metabolic pathway.
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影响因子：
4.2
作者：
Sempere, Angela;Arias, Angela;Campistol, Jaume
通讯作者：
Campistol, Jaume
影响因子：
64.5
作者：
Narkar VA;Downes M;Yu RT;Embler E;Wang YX;Banayo E;Mihaylova MM;Nelson MC;Zou Y;Juguilon H;Kang H;Shaw RJ;Evans RM
通讯作者：
Evans RM
影响因子：
2.9
作者：
MEYER, E;LEONARD, NJ;SMITH, JM
通讯作者：
SMITH, JM
影响因子：
4.8
作者：
Rattan, R;Giri, S;Singh, I
通讯作者：
Singh, I
影响因子：
4.1
作者：
Becker, Jennifer;Hackl, Matthias;Brinkrolf, Karina
通讯作者：
Brinkrolf, Karina