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首页> 外文期刊>Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society >ARNO but not cytohesin-1 translocation is phosphatidylinositol 3-kinase-dependent in HL-60 cells.
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ARNO but not cytohesin-1 translocation is phosphatidylinositol 3-kinase-dependent in HL-60 cells.

机译：在HL-60细胞中，ARNO而不是细胞粘附素1易位是磷脂酰肌醇3激酶依赖性的。

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摘要

Cytohesin-1 and ARNO are guanine nucleotide-exchange factors (GEFs) for ADP-ribosylation factor (Arf). Here, we show that ARNO is expressed in HL-60 cells and established that granulocytic differentiation induced with Me2SO stimulated cytohesin-1 but not ARNO expression. Cytohesin-1 levels in HL-60 granulocytes were similar to those in human neutrophils. Me2SO-differentiated HL-60 cells expressed ARNO and cytohesin-1 isoforms with a diglycine and a triglycine motif in their PH domains, respectively. In vitro, ARNO diglycine and cytohesin-1 triglycine enhanced phospholipase D1 (PLD1) activation by Arf1 with near-maximal effects at 250 nM. These effects were marked particularly at low Mg2+ concentrations. PLD activation was well-correlated with GTP binding to Arf1, and cytohesin-1 was always more potent than ARNO in the PLD- and GTP-binding assays. Increasing Mg2+ concentrations reduced PLD and Arf1 activation by Arf-GEFs. fMetLeuPhe and phorbol 12-myristate 13-acetate stimulated ARNO and cytohesin-1 as well as Arf1 translocation to HL-60 cell membranes. fMetLeuPhe-mediated ARNO recruitment, but not cytohesin-1 and Arf1 translocation, was blocked by phosphatidylinositol 3-kinase inhibitors. The combined results demonstrate that cytohesin-1 triglycine participates in a major phosphatidylinositol 3-kinase-independent pathway linking cell-surface receptors to Arf1 activation and translocation in human granulocytes.
机译：Cytohesin-1和ARNO是ADP-核糖基化因子（Arf）的鸟嘌呤核苷酸交换因子（GEF）。在这里，我们显示ARNO在HL-60细胞中表达，并建立了用Me2SO诱导的粒细胞分化刺激了细胞粘附素1，但不刺激ARNO表达。 HL-60粒细胞中的细胞黏着蛋白1水平与人中性粒细胞中的相似。 Me2SO分化的HL-60细胞表达在其PH域中分别具有双甘氨酸和三甘氨酸基序的ARNO和细胞粘附素1亚型。在体外，ARNO双甘氨酸和cytohesin-1三甘氨酸增强了Arf1对磷脂酶D1（PLD1）的激活，在250 nM时具有接近最大的作用。这些影响在低Mg2 +浓度下尤为明显。 PLD激活与GTP与Arf1的结合密切相关，并且在PLD和GTP结合测定中，cytohesin-1总是比ARNO更有效。 Mg2 +浓度的增加会降低Arf-GEF的PLD和Arf1激活。 fMetLeuPhe和佛波醇12-肉豆蔻酸酯13-乙酸酯刺激了ARNO和cytohesin-1以及Arf1易位至HL-60细胞膜。 fMetLeuPhe介导的ARNO募集被磷脂酰肌醇3激酶抑制剂阻滞，但不使细胞黏附素1和Arf1易位。合并的结果表明，cytohesin-1 triglycine参与了主要的磷脂酰肌醇3激酶非依赖性途径，该途径将细胞表面受体与人粒细胞中的Arf1活化和易位联系起来。

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