Adenylate kinase isozyme 2 is essential for growth and development of Drosophila melanogaster
Adenylate kinase isozyme 2 is essential for growth and development of Drosophila melanogaster
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DOI：
10.1016/j.cbpb.2009.01.006
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发表时间：
2009-05-01
影响因子：
2.2
通讯作者：
Noma, Takafumi
中科院分区：
文献类型：
--
作者：
Fujisawa, Koichi;Murakami, Ryutaro;Noma, Takafumi
腺苷酸激酶在系统发育上广泛存在、高度保守，并参与能量代谢和能量转移。其中，腺苷酸激酶 (AK) 同工酶 2 独特地定位于线粒体膜间隙，其生理作用仍然很大程度上未知。在本研究中，我们选择了黑腹果蝇来分析其在体内的作用。克隆了 AK 同工酶 cDNA，并在黑腹果蝇中表征了它们的基因表达。推导的氨基酸序列包含高度保守的 P 环基序、NMP 结合基序和 AK 的 LID 结构域。此外，利用P-element技术检查了AK2基因敲除对AK2突变体表型的影响。尽管纯合 AK2 突变胚胎发育时没有任何可见缺陷，但它们的生长停止并在到达第三龄幼虫阶段之前死亡。在受精卵中检测到母体提供的 AI(2 mRNA)，并且在纯合 AK2 突变体的一龄和二龄幼虫中观察到弱 AK2 活性，表明母体提供的 AK2 足以满足胚胎发育。幼虫阶段 AK2 活性的消失导致生长停滞并最终死亡。这些结果表明 AK2 在腺嘌呤核苷酸代谢中发挥着关键作用 存在于线粒体膜间隙中，对于黑腹果蝇的生长至关重要。 (C) 2009 Elsevier Inc. 保留所有权利。
Adenylate kinases are phylogenetically widespread, highly conserved, and involved in energy metabolism and energy transfer Of these, adenylate kinase (AK) isozyme 2 is uniquely localized in the mitochondrial intermembrane space and its physiological role remains largely unknown. In this study, we selected Drosophila melanogaster to analyze its role in vivo. AK isozyme cDNAs were cloned and their gene expressions were characterized in D. melanogaster. The deduced amino acid sequences contain highly conserved motifs for P-loop, NMP binding, and LID domains of AKs. In addition, the effects of AK2 gene knockout on phenotype of AK2 mutants were examined using P-element technology. Although homozygous AK2 mutated embryos developed without any visible defects, their growth ceased and they died before reaching the third instar larval stage. Maternally provided AI(2 mRNA was detected in fertilized eggs, and weak AK2 activity was observed in first and second instar larvae of the homozygous AK2 mutants, suggesting that maternally provided AK2 is sufficient for embryonic development. Disappearance of AK2 activity during larval stages resulted in growth arrest and eventual death. These results demonstrate that AK2 plays a critical role in adenine nucleotide metabolism in the mitochondrial intermembrane space and is essential for growth in D. melanogaster. (C) 2009 Elsevier Inc. All rights reserved.