Apical expression of three RhopH1/Clag proteins as components of the Plasmodium falciparum RhopH complex
Apical expression of three RhopH1/Clag proteins as components of the Plasmodium falciparum RhopH complex
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DOI：
10.1016/j.molbiopara.2005.05.003
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发表时间：
2005-09-01
影响因子：
1.5
通讯作者：
Torii, M
中科院分区：
文献类型：
--
作者：
Kaneko, O;Lim, BYSY;Torii, M
恶性疟原虫高分子量棒状体蛋白（PfRhopH）复合物对于寄生虫生长是重要的，并且包括三种不同的基因产物：RhopH 1、RhopH 2和RhopH 3。我们以前已经表明，恶性疟原虫RhopH 1是由PFC 0110 w（clag3.2）或PFC 0120 w（clag3.1），以前命名的clag（细胞粘附连锁无性基因）多基因家族的成员编码。在这份报告中，我们进一步的特点rhoph 1/clag成员的基因结构，转录和蛋白表达。确定了所有5个rhoph 1/clag成员的cDNA序列，证实了先前对内含子-外显子边界的计算机预测。所有成员基因在HB 3和3D 7中均有转录，但clag3.2在Dd 2中不转录。所有基因的转录本的峰值丰度观察到在后期的启动子阶段。Clag 2和Clag3.1的特异性抗血清将这些蛋白质定位于裂殖子的顶端，发现这两种蛋白质都是PfRhopH复合物的组分。用抗Clag 9抗体免疫沉淀的PfRhopH复合物既不含Clag 2也不含Clag3.1，从而表明PfRhopH复合物仅含有单独的rhoph 1/clag基因产物。由于PfRhopH复合物结合红细胞表面，并且RhopH 2和RhopH 3由单拷贝基因编码，因此RhopH 1/Clag蛋白可以用于赋予个体复合物的作用一定程度的特异性。(c)2005 Elsevier B. V.保留所有权利。
The Plasmodium falciparum high molecular mass rhoptry protein (`PfRhopH') complex is important for parasite growth and comprises three distinct gene products: RhopH1, RhopH2 and RhopH3. We have previously shown that P. falciparum RhopH1 is encoded by either PFC0110w (clag3.2) or PFC0120w (clag3.1), members of the previously-named clag (cytoadherence-linked asexual gene) multigene family. In this report, we have further characterized rhoph1/clag members in terms of gene structure, transcription and protein expression. The cDNA sequences for all five rhoph1/clag members were determined, confirming previous in silico predictions of intron-exon boundaries. All member genes were transcribed in HB3 and 3D7 parasite lines, but clag3.2 was not transcribed in Dd2 parasites. The peak abundance of transcripts for all genes was observed during the late schizont stage. Antisera specific to Clag2 and Clag3.1 localized these proteins to the apical end of merozoites in segmented schizonts, and both proteins are found to be components of the PfRhopH complex. PfRhopH complex that was immunoprecipitated with anti-Clag9 antibody contained neither Clag2 nor Clag3.1, thereby suggesting that PfRhopH complexes contain only individual rhoph1/clag gene products. Since the PfRhopH complex binds the erythrocyte surface, and RhopH2 and RhopH3 are encoded by single copy genes, the RhopH1/Clag proteins may serve to confer some degree of specificity to the roles of the individual complexes. (c) 2005 Elsevier B.V. All rights reserved.