Inflammation Biology Section, Laboratory of Molecular Immunology, NIAID, National Institutes of Health, Bethesda, Maryland 20892, USA.
J Biol Chem. 2010 Nov 12;285(46):35537-50. doi: 10.1074/jbc.M110.128033. Epub 2010 Sep 9.
Because T cell differentiation leads to an expanded repertoire of chemokine receptors, a subgroup of G protein-coupled receptors, we hypothesized that the repertoire of G proteins might be altered in parallel. We analyzed the abundance of mRNA and/or protein of six G protein α-subunits in human CD4(+) and CD8(+) T cell subsets from blood. Although most G protein α-subunits were similarly expressed in all subsets, the abundance of Gα(o), a protein not previously described in hematopoietic cells, was much higher in memory versus naive cells. Consistent with these data, activation of naive CD4(+) T cells in vitro significantly increased the abundance of Gα(o) in cells stimulated under nonpolarizing or T(H)17 (but not T(H)1 or T(H)2)-polarizing conditions. In functional studies, the use of a chimeric G protein α-subunit, Gα(qo5), demonstrated that chemokine receptors could couple to Gα(o)-containing G proteins. We also found that Gα(i1), another α-subunit not described previously in leukocytes, was expressed in naive T cells but virtually absent from memory subsets. Corresponding to their patterns of expression, siRNA-mediated knockdown of Gα(o) in memory (but not naive) and Gα(i1) in naive (but not memory) CD4(+) T cells inhibited chemokine-dependent migration. Moreover, although even in Gα(o)- and Gα(i1)-expressing cells mRNAs of these α-subunits were much less abundant than Gα(i2) or Gα(i3), knockdown of any of these subunits impaired chemokine receptor-mediated migration similarly. Together, our data reveal a change in the repertoire of Gα(i/o) subunits during T cell differentiation and suggest functional equivalence among Gα(i/o) subunits irrespective of their relative abundance.
由于 T 细胞分化导致趋化因子受体(一组 G 蛋白偶联受体)谱扩大,我们假设 G 蛋白谱可能会平行改变。我们分析了来自血液的人 CD4(+)和 CD8(+)T 细胞亚群中六种 G 蛋白α-亚基的 mRNA 和/或蛋白丰度。虽然大多数 G 蛋白α-亚基在所有亚群中表达相似,但以前在造血细胞中未描述的 Gα(o)的丰度在记忆细胞中明显高于幼稚细胞。与这些数据一致,体外激活幼稚 CD4(+)T 细胞可显著增加在非极化或 T(H)17(而非 T(H)1 或 T(H)2)极化条件下刺激的细胞中 Gα(o)的丰度。在功能研究中,使用嵌合 G 蛋白α-亚基 Gα(qo5)表明趋化因子受体可以与含有 Gα(o)的 G 蛋白偶联。我们还发现,另一种以前未在白细胞中描述的 Gα(i1)α-亚基在幼稚 T 细胞中表达,但在记忆亚群中几乎不存在。与其表达模式相对应,siRNA 介导的记忆(而非幼稚)中的 Gα(o)和幼稚(而非记忆)中的 Gα(i1)敲低抑制了趋化因子依赖性迁移。此外,尽管即使在表达 Gα(o)和 Gα(i1)的细胞中,这些α-亚基的 mRNA 丰度也远低于 Gα(i2)或 Gα(i3),但敲低任何这些亚基都同样损害了趋化因子受体介导的迁移。总之,我们的数据揭示了 T 细胞分化过程中 Gα(i/o)亚基谱的变化,并表明 Gα(i/o)亚基之间存在功能等同性,而与它们的相对丰度无关。