McKeveney P J, Hodges V M, Mullan R N, Maxwell P, Simpson D, Thompson A, Winter P C, Lappin T R, Maxwell A P
Department of Nephrology, Belfast City Hospital, The Queen's University of Belfast, Tower Block, Lisburn Road, Belfast BT9 7AB, Northern Ireland, UK.
Br J Haematol. 2001 Mar;112(4):1016-24. doi: 10.1046/j.1365-2141.2001.02644.x.
Gene expression profiles during erythropoietin (Epo)-induced differentiation of erythroid progenitor cells derived from the Friend virus anaemia (FVA) and phenylhydrazine (PHZ) murine models have been examined using differential display polymerase chain reaction (PCR). Ten cDNA fragments upregulated by Epo were isolated. The ribonuclease protection assay confirmed differential expression between Epo-stimulated and Epo-deprived cells for one of these, provisionally named ERIC-1. Sequencing of the full-length cDNA predicted a protein of 558 amino acids, 17 amino acids longer than mTACC3, the third member of a novel family of proteins that contain a coiled-coil domain. The human homologue, cloned using rapid amplification of cDNA ends (RACE)-PCR, encodes a larger protein of 838 amino acids that is identical to hTACC3. In addition to erythroid precursor cells, ERIC-1/TACC3 is expressed at high levels in the testes, at moderate levels in the thymus and peripheral leucocytes, and at lower levels in the spleen and intestinal tissue. Immunohistochemical analysis using an antibody to a GST fusion product of the C-terminus of hERIC-1/TACC3 revealed that it is localized to Sertoli cells in the human testes. Confocal microscopy demonstrated hERIC-1/TACC3 protein concentrated in the perinuclear vesicles of dermal microvascular endothelial cells. Although ERIC-1/TACC3 is expressed in a wide range of tissues, its upregulation by Epo in erythroid progenitors implies that it has a role in terminal erythropoiesis.
利用差异显示聚合酶链反应(PCR)检测了促红细胞生成素(Epo)诱导的源自弗氏病毒贫血(FVA)和苯肼(PHZ)小鼠模型的红系祖细胞分化过程中的基因表达谱。分离出了10个受Epo上调的cDNA片段。核糖核酸酶保护试验证实了其中一个(暂命名为ERIC-1)在Epo刺激细胞和Epo缺失细胞之间存在差异表达。全长cDNA测序预测出一种含558个氨基酸的蛋白质,比mTACC3长17个氨基酸,mTACC3是一个包含卷曲螺旋结构域的新蛋白质家族的第三个成员。使用cDNA末端快速扩增(RACE)-PCR克隆的人类同源物编码一种更大的含838个氨基酸的蛋白质,与hTACC3相同。除了红系前体细胞外,ERIC-1/TACC3在睾丸中高水平表达,在胸腺和外周白细胞中中等水平表达,在脾脏和肠道组织中低水平表达。使用针对hERIC-1/TACC3 C末端的GST融合产物的抗体进行免疫组织化学分析显示,它定位于人类睾丸的支持细胞中。共聚焦显微镜显示hERIC-1/TACC3蛋白集中在真皮微血管内皮细胞的核周小泡中。尽管ERIC-1/TACC3在多种组织中表达,但其在红系祖细胞中被Epo上调表明它在终末红细胞生成中起作用。