Wu Jui-Ching, Rose Lesilee S
Section of Molecular and Cellular Biology, University of California, Davis, CA 95616, USA.
Mol Biol Cell. 2007 Nov;18(11):4470-82. doi: 10.1091/mbc.e07-02-0105. Epub 2007 Aug 29.
The conserved PAR proteins are localized in asymmetric cortical domains and are required for the polarized localization of cell fate determinants in many organisms. In Caenorhabditis elegans embryos, LET-99 and G protein signaling act downstream of the PARs to regulate spindle positioning and ensure asymmetric division. PAR-3 and PAR-2 localize LET-99 to a posterior cortical band through an unknown mechanism. Here we report that LET-99 asymmetry depends on cortically localized PAR-1 and PAR-4 but not on cytoplasmic polarity effectors. In par-1 and par-4 embryos, LET-99 accumulates at the entire posterior cortex, but remains at low levels at the anterior cortex occupied by PAR-3. Further, PAR-3 and PAR-1 have graded cortical distributions with the highest levels at the anterior and posterior poles, respectively, and the lowest levels of these proteins correlate with high LET-99 accumulation. These results suggest that PAR-3 and PAR-1 inhibit the localization of LET-99 to generate a band pattern. In addition, PAR-1 kinase activity is required for the inhibition of LET-99 localization, and PAR-1 associates with LET-99. Finally, examination of par-1 embryos suggests that the banded pattern of LET-99 is critical for normal posterior spindle displacement and to prevent spindle misorientation caused by cell shape constraints.
保守的PAR蛋白定位于不对称的皮层结构域,是许多生物体中细胞命运决定因子极化定位所必需的。在秀丽隐杆线虫胚胎中,LET-99和G蛋白信号在PAR蛋白下游起作用,以调节纺锤体定位并确保不对称分裂。PAR-3和PAR-2通过未知机制将LET-99定位于后皮层带。在此,我们报道LET-99的不对称性取决于皮层定位的PAR-1和PAR-4,而不依赖于细胞质极性效应器。在par-1和par-4胚胎中,LET-99在后皮层整体积累,但在被PAR-3占据的前皮层中仍保持低水平。此外,PAR-3和PAR-1在皮层呈梯度分布,分别在前极和后极水平最高,这些蛋白的最低水平与LET-99的高积累相关。这些结果表明,PAR-3和PAR-1抑制LET-99的定位以产生带状模式。此外,PAR-1激酶活性是抑制LET-99定位所必需的,且PAR-1与LET-99相关联。最后,对par-1胚胎的检查表明,LET-99的带状模式对于正常的后纺锤体移位以及防止由细胞形状限制引起的纺锤体错误定向至关重要。