Karlsson Susann, Kowanetz Katarzyna, Sandin Asa, Persson Camilla, Ostman Arne, Heldin Carl-Henrik, Hellberg Carina
Ludwig Institute for Cancer Research, Uppsala University Biomedical Center, S-751 24 Uppsala, Sweden.
Mol Biol Cell. 2006 Nov;17(11):4846-55. doi: 10.1091/mbc.e06-04-0306. Epub 2006 Sep 13.
We have previously shown that the T-cell protein tyrosine phosphatase (TC-PTP) dephosphorylates the platelet-derived growth factor (PDGF) beta-receptor. Here, we show that the increased PDGF beta-receptor phosphorylation in TC-PTP knockout (ko) mouse embryonic fibroblasts (MEFs) occurs primarily on the cell surface. The increased phosphorylation is accompanied by a TC-PTP-dependent, monensin-sensitive delay in clearance of cell surface PDGF beta-receptors and delayed receptor degradation, suggesting PDGF beta-receptor recycling. Recycled receptors could also be directly detected on the cell surface of TC-PTP ko MEFs. The effect of TC-PTP depletion was specific for the PDGF beta-receptor, because PDGF alpha-receptor homodimers were cleared from the cell surface at the same rate in TC-PTP ko MEFs as in wild-type MEFs. Interestingly, PDGF alphabeta-receptor heterodimers were recycling. Analysis by confocal microscopy revealed that, in TC-PTP ko MEFs, activated PDGF beta-receptors colocalized with Rab4a, a marker for rapid recycling. In accordance with this, transient expression of a dominant-negative Rab4a construct increased the rate of clearance of cell surface receptors on TC-PTP ko MEFs. Thus, loss of TC-PTP specifically redirects the PDGF beta-receptor toward rapid recycling, which is the first evidence of differential trafficking of PDGF receptor family members.
我们之前已经表明,T细胞蛋白酪氨酸磷酸酶(TC-PTP)可使血小板衍生生长因子(PDGF)β受体去磷酸化。在此,我们发现,在TC-PTP基因敲除(ko)小鼠胚胎成纤维细胞(MEF)中,PDGFβ受体磷酸化增加主要发生在细胞表面。磷酸化增加伴随着细胞表面PDGFβ受体清除过程中依赖于TC-PTP且对莫能菌素敏感的延迟以及受体降解延迟,提示存在PDGFβ受体循环利用。在TC-PTP基因敲除的MEF细胞表面也能直接检测到循环利用的受体。TC-PTP缺失的影响对PDGFβ受体具有特异性,因为在TC-PTP基因敲除的MEF中,PDGFα受体同二聚体从细胞表面清除的速率与野生型MEF相同。有趣的是,PDGFαβ受体异二聚体存在循环利用现象。共聚焦显微镜分析显示,在TC-PTP基因敲除的MEF中,活化的PDGFβ受体与Rab4a共定位,Rab4a是快速循环利用的标志物。与此一致的是,显性负性Rab4a构建体的瞬时表达增加了TC-PTP基因敲除的MEF细胞表面受体的清除速率。因此,TC-PTP的缺失特异性地使PDGFβ受体转向快速循环利用,这是PDGF受体家族成员差异性运输的首个证据。