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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Membrane-associated and soluble granulocyte/macrophage-colony-stimulating factor receptor alpha subunits are independently regulated in HL-60 cells.
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Membrane-associated and soluble granulocyte/macrophage-colony-stimulating factor receptor alpha subunits are independently regulated in HL-60 cells.

机译：膜相关和可溶性粒细胞/巨噬细胞集落刺激因子受体α亚基在HL-60细胞中受到独立调节。

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摘要

The effects of granulocyte/macrophage-colony-stimulating factor (GM-CSF) are mediated by interaction with its composite receptor (GMR), which consists of a unique alpha subunit (GMR alpha) and a beta subunit (GMR beta) that is common to the receptors for GM-CSF, interleukin 3, and interleukin 5. GMR beta is required for high-affinity binding, cell proliferation, and protein phosphorylation but has no intrinsic GM-CSF-binding activity. GMR alpha in isolation binds to GM-CSF with low affinity and can signal for increased glucose uptake. In addition to the membrane-bound receptor (mGMR alpha), there is a naturally occurring soluble isoform (sGMR alpha) that is released free into the pericellular milieu. Analysis of genomic sequences reveals that the soluble GMR alpha isoform comes about by alternative mRNA splicing. To examine GMR alpha expression, we developed a quantitative reverse transcription-polymerase chain reaction assay based on serial dilutions of in vitro transcribed GMR alpha RNA. This assay provides a strict log-log measure of GMR alpha RNA expression, distinguishes transcripts related to the soluble and membrane-associated isoforms, and quantitatively detects 0.1 fg of GMR alpha-related mRNA. There was little or no GMR alpha expression in two human lymphoid cell lines and in the erythroblastic leukemia cell line K562, but all myeloid cell lines tested expressed both the membrane-associated and soluble isoforms of GMR alpha. Baseline level of expression of both isoforms varied > 20-fold among the myeloid cell lines studied. Differentiation of HL-60 cells to neutrophils with dimethyl sulfoxide led to a 2-fold downregulation of sGMR alpha and a 20-fold upregulation of mGMR alpha. These differentiation-induced transcriptional changes were unrelated to changes in mRNA stability. These findings indicate that sGMR alpha is differentially expressed from mGMR alpha in human hematopoietic cells and that programmed downregulation of sGMR alpha may be important in myeloid maturation.
机译：粒细胞/巨噬细胞集落刺激因子（GM-CSF）的作用是通过与其复合受体（GMR）相互作用而介导的，复合受体由独特的α亚基（GMR alpha）和常见的β亚基（GMR beta）组成GM-CSF，白介素3和白介素5的受体具有GMRβ。它需要高亲和力结合，细胞增殖和蛋白质磷酸化，但不具有固有的GM-CSF结合活性。单独的GMRα以低亲和力与GM-CSF结合，并可以发出增加葡萄糖摄取的信号。除了膜结合受体（mGMRα）外，还有一种天然存在的可溶性同工型（sGMRα），可以自由释放到细胞周围环境中。对基因组序列的分析表明，可溶性GMRα亚型是通过替代性的mRNA剪接产生的。为了检查GMRα的表达，我们基于体外转录的GMRαRNA的系列稀释液开发了定量逆转录聚合酶链反应法。该测定法提供了对GMRαRNA表达的严格对数对数测量，区分了与可溶性和膜相关同工型有关的转录本，并定量检测了0.1 fg的GMRα相关mRNA。在两个人淋巴样细胞系和成幼红细胞白血病细胞系K562中几乎没有GMRα表达，但是所有测试的髓样细胞系均表达了GMRα的膜相关和可溶性同工型。在研究的髓样细胞系中，两种同工型的基线表达水平变化> 20倍。用二甲基亚砜将HL-60细胞分化为嗜中性粒细胞，导致sGMRα上调2倍，而mGMRα上调20倍。这些分化诱导的转录变化与mRNA稳定性的变化无关。这些发现表明在人类造血细胞中sGMR alpha与mGMR alpha差异表达，并且sGMR alpha的程序下调在骨髓成熟中可能很重要。

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