GPR125 (ADGRA3) is an autocleavable adhesion GPCR that traffics with Dlg1 to the basolateral membrane and regulates epithelial apicobasal polarity.
GPR125 (ADGRA3) is an autocleavable adhesion GPCR that traffics with Dlg1 to the basolateral membrane and regulates epithelial apicobasal polarity.
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DOI：
10.1016/j.jbc.2022.102475
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发表时间：
2022-10
影响因子：
4.8
通讯作者：
Sumimoto, Hideki
中科院分区：
文献类型：
--
作者：
Sakurai, Tsubasa;Kamakura, Sachiko;Hayase, Junya;Kohda, Akira;Nakamura, Masafumi;Sumimoto, Hideki
G蛋白偶联受体（GPCR）的粘附家族由N-末端大的胞外区定义，所述胞外区包含各种粘附相关结构域和高度保守的GPCR-自蛋白水解诱导（GAIN）结构域，后者位于紧接典型的七跨膜结构域之前。这些受体广泛表达并参与多种功能，包括发育、血管生成、突触形成和肿瘤发生。GPR 125（ADGRA 3），孤儿粘附GPCR，已被证明可以调节平面细胞极性在原肠胚斑马鱼，但其生化特性和在哺乳动物细胞中的作用仍然是未知的。在这里，我们表明，人GPR 125可能经历顺式自蛋白水解时，在犬肾上皮MDCK细胞和人胚肾HEK 293细胞表达。切割似乎发生在一个非典型的GPCR蛋白水解位点内的GAIN域在受体生物合成的早期阶段。产品，即，N-末端和C-末端片段，似乎在自身蛋白水解后保持结合，如在其它粘附GPCR中观察到的。此外，在极化的MDCK细胞中，GPR 125仅被募集到质膜的基底外侧域。募集可能需要GPR 125的C末端PDZ结构域结合基序及其与细胞极性蛋白Dlg 1的相互作用。GPR 125以及Dlg 1的敲低导致在MDCK细胞的Matrigel 3D培养物中形成具有多个腔的异常囊肿。与多腔表型一致，在GPR 125-KO MDCK细胞中，有丝分裂纺锤体在囊形成期间方向不正确。因此，基底外侧蛋白GPR 125，一种可自切割的粘附GPCR，似乎在上皮细胞的顶基底极化中起着至关重要的作用。
The adhesion family of G protein–coupled receptors (GPCRs) is defined by an N-terminal large extracellular region that contains various adhesion-related domains and a highly-conserved GPCR-autoproteolysis-inducing (GAIN) domain, the latter of which is located immediately before a canonical seven-transmembrane domain. These receptors are expressed widely and involved in various functions including development, angiogenesis, synapse formation, and tumorigenesis. GPR125 (ADGRA3), an orphan adhesion GPCR, has been shown to modulate planar cell polarity in gastrulating zebrafish, but its biochemical properties and role in mammalian cells have remained largely unknown. Here, we show that human GPR125 likely undergoes cis-autoproteolysis when expressed in canine kidney epithelial MDCK cells and human embryonic kidney HEK293 cells. The cleavage appears to occur at an atypical GPCR proteolysis site within the GAIN domain during an early stage of receptor biosynthesis. The products, i.e., the N-terminal and C-terminal fragments, seem to remain associated after self-proteolysis, as observed in other adhesion GPCRs. Furthermore, in polarized MDCK cells, GPR125 is exclusively recruited to the basolateral domain of the plasma membrane. The recruitment likely requires the C-terminal PDZ-domain–binding motif of GPR125 and its interaction with the cell polarity protein Dlg1. Knockdown of GPR125 as well as that of Dlg1 results in formation of aberrant cysts with multiple lumens in Matrigel 3D culture of MDCK cells. Consistent with the multilumen phenotype, mitotic spindles are incorrectly oriented during cystogenesis in GPR125-KO MDCK cells. Thus, the basolateral protein GPR125, an autocleavable adhesion GPCR, appears to play a crucial role in apicobasal polarization in epithelial cells.
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