Eimeria maxima phosphatidylinositol 4-phosphate 5-kinase: locus sequencing, characterization, and cross-phylum comparison.
Eimeria maxima phosphatidylinositol 4-phosphate 5-kinase: locus sequencing, characterization, and cross-phylum comparison.
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艾美耳球虫磷脂酰肌醇 4-磷酸 5-激酶：基因座测序、表征和跨门比较。
DOI：
10.1007/s00436-010-2104-7
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发表时间：
2011
影响因子：
2
通讯作者：
Goh MY
中科院分区：
文献类型：
--
作者：
Goh MY
磷脂酰肌醇 4-磷酸 5-激酶 (PIP5K) 可能在艾美球虫属原生动物寄生虫入侵宿主细胞中发挥重要作用，这种寄生虫可导致牲畜严重肠道疾病。在这里，我们报道了极大艾美耳球虫（Weybridge 株）中 PIP5K 基因的结构组织。二E．携带E的maximaBAC克隆。选择 maxima PIP5K(EmPIP5K) 编码序列进行鸟枪法测序，产生 9.1 kb 的基因组片段。 EmPIP5K 编码区最初是使用计算机基因预测方法进行鉴定的，随后通过将 cDNA 末端的快速扩增和 RT-PCR 生成的 cDNA 序列映射到其基因组片段来确认。推定的EmPIP5K基因位于互补链上的710-8036nt位置并且由23个外显子组成。将 1147 个氨基酸序列与先前注释的来自其他 Apicomplexa 物种的 PIP5K 蛋白进行比对，检测到包含激酶核心结构域的三个保守基序，先前的蛋白缺失研究已表明这对于 PIP5K 蛋白功能是必需的。系统发育分析进一步证明推定的EmPIP5K 蛋白与其他Apicomplexa 蛋白是直系同源的。随后的比较基因结构表征揭示了 apicomplexanPIP5K 基因整个进化过程中内含子丢失/获得的事件。对基因组结构的进一步检查揭示了两个基序区域之间可能存在“内含子增益”的趋势。我们的研究结果提供了对 PIP5K 基因座进化过程中发生的结构变异的初步见解，并可能有助于了解该基因在顶复门寄生虫的细胞生物学中的功能作用。
Phosphatidylinositol 4-phosphate 5-kinase (PIP5K) may play an important role in host-cell invasion by theEimeriaspecies, protozoan parasites which can cause severe intestinal disease in livestock. Here, we report the structural organization of thePIP5Kgene inEimeria maxima(Weybridge strain). TwoE. maximaBAC clones carrying theE. maxima PIP5K(EmPIP5K) coding sequences were selected for shotgun sequencing, yielding a 9.1-kb genomic segment. TheEmPIP5Kcoding region was initially identified using in silico gene-prediction approaches and subsequently confirmed by mapping rapid amplification of cDNA ends and RT-PCR-generated cDNA sequence to its genomic segment. The putativeEmPIP5Kgene was located at position 710-8036 nt on the complimentary strand and comprised of 23 exons. Alignment of the 1147 amino acid sequence with previously annotated PIP5K proteins from other Apicomplexa species detected three conserved motifs encompassing the kinase core domain, which has been shown by previous protein deletion studies to be necessary for PIP5K protein function. Phylogenetic analysis provided further evidence that the putativeEmPIP5K protein is orthologous to that of other Apicomplexa. Subsequent comparative gene structure characterization revealed events of intron loss/gain throughout the evolution of the apicomplexanPIP5Kgene. Further scrutiny of the genomic structure revealed a possible trend towards “intron gain” between two of the motif regions. Our findings offer preliminary insights into the structural variations that have occurred during the evolution of thePIP5Klocus and may aid in understanding the functional role of this gene in the cellular biology of apicomplexan parasites.