INTERLEUKIN-3 FACILITATES GLUCOSE-TRANSPORT IN A MYELOID CELL-LINE BY REGULATING THE AFFINITY OF THE GLUCOSE-TRANSPORTER FOR GLUCOSE - INVOLVEMENT OF PROTEIN-PHOSPHORYLATION IN TRANSPORTER ACTIVATION
INTERLEUKIN-3 FACILITATES GLUCOSE-TRANSPORT IN A MYELOID CELL-LINE BY REGULATING THE AFFINITY OF THE GLUCOSE-TRANSPORTER FOR GLUCOSE - INVOLVEMENT OF PROTEIN-PHOSPHORYLATION IN TRANSPORTER ACTIVATION
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DOI：
10.1042/bj3050843
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发表时间：
1995-02-01
影响因子：
4.1
通讯作者：
TAN, AS
中科院分区：
文献类型：
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作者：
BERRIDGE, MV;TAN, AS
生长因子通过激活导致细胞周期进展和差异基因表达的信号转导途径来促进细胞存活和增殖。基础细胞代谢和细胞生长和增殖所必需的大分子合成所需的单糖的摄取被认为是信号转导到细胞核的结果。然而，在存在DNA合成和呼吸抑制剂的情况下，生长因子仍然可以在短期内促进细胞存活反应，这提高了它们也可以调节细胞存活所必需的关键膜和胞质过程的可能性。我们通过研究造血生长因子白细胞介素-3（IL-3）在骨髓源性细胞系32 D葡萄糖转运调节中的作用，直接验证了这一假设。我们发现IL-3通过积极维持质膜葡萄糖转运蛋白对葡萄糖的亲和力来促进葡萄糖转运（K-m 1.35+/-0.15 mM，n = 4）。停止IL-3 1小时导致对葡萄糖的亲和力降低（K-m 2.96+/-0.28 mM，n = 4），而V-max无相关变化。此外，细胞松弛素B结合到分离的质膜，在细胞表面的葡萄糖转运蛋白分子，没有显着差异之间的控制和IL-3处理的细胞。用丝裂霉素C或用呼吸毒物叠氮化钠抑制DNA合成，并不影响IL-3促进葡萄糖转运的能力。相反，酪氨酸激酶抑制剂染料木素和erbstatin广泛抑制控制和IL-3刺激的葡萄糖转运，在低抑制剂浓度下观察到IL-3刺激的反应的一些偏好。光激活蛋白激酶C抑制剂calphostin C也抑制对照和IL-3刺激的葡萄糖转运，但对IL-3反应没有偏好。此外，酪氨酸磷酸酶抑制剂原钒酸盐刺激对照和IL-3依赖性葡萄糖转运50-80%，而蛋白激酶A抑制剂KT 5720在平台值抑制葡萄糖转运约20%。这些结果表明，IL-3参与葡萄糖转运蛋白活性的持续维持的机制，涉及酪氨酸激酶和蛋白激酶C，并证明这种激活是不依赖于呼吸或信号转导到细胞核。
Growth factors promote cell survival and proliferation by activating signal transduction pathways that result in progression through the cell cycle and differential gene expression. Uptake of simple sugars needed for basal cell metabolism, and for macromolecular synthesis necessary for cell growth and proliferation, is thought to follow as a consequence of signal transduction to the nucleus. However, in the presence of inhibitors of DNA synthesis and respiration, growth factors can still promote cell survival responses in the short term, raising the possibility that they may also regulate critical membrane and cytosolic processes necessary for cell survival. We have tested this hypothesis directly by investigating the role of the haemopoietic growth factor, interIeukin-3 (IL-3), in the regulation of glucose transport in the bone marrow-derived cell line, 32D. We show that IL-3 promotes glucose transport by actively maintaining the affinity of the plasma membrane glucose transporter for glucose (K-m 1.35+/-0.15 mM, n = 4). Withdrawal of IL-3 for 1 h resulted in reduced affinity for glucose (K-m 2.96+/-0.28 mM, n = 4) without an associated change in V-max. Furthermore, glucose transporter molecules at the cell surface, as determined by cytochalasin B binding to isolated plasma membranes, did not differ significantly between control and IL-3-treated cells. Inhibition of DNA synthesis with mitomycin C or with the respiratory poison, sodium azide, did not affect the ability of IL-3 to promote glucose transport. In contrast, the tyrosine kinase inhibitors genistein and erbstatin extensively inhibited control and IL-3-stimulated glucose transport, some preference for IL-3-stimulated responses being observed at low inhibitor concentrations. The light-activated protein kinase C inhibitor, calphostin C, also inhibited control and IL-3-stimulated glucose transport but without preference for IL-3 responses. Additionally, the tyrosine phosphatase inhibitor, orthovanadate, stimulated control and IL-3-dependent glucose transport by 50-80% while the protein kinase A inhibitor, KT5720, inhibited glucose transport by about 20% at plateau values. These results indicate that IL-3 is involved in continuous maintenance of glucose transporter activity by a mechanism that involves tyrosine kinases and protein kinase C, and demonstrate that this activation is not dependent on respiration or signal transduction to the nucleus.