Department of Biochemistry, University of Oxford , South Parks Road, Oxford OX1 3QU , UK.
Biol Open. 2013 Jun 6;2(7):718-27. doi: 10.1242/bio.20134549. Print 2013 Jul 15.
Caenorhabditis elegans seam cells divide in the stem-like mode throughout larval development, with the ability to both self-renew and produce daughters that differentiate. Seam cells typically divide asymmetrically, giving rise to an anterior daughter that fuses with the hypodermis and a posterior daughter that proliferates further. Previously we have identified rnt-1 (a homologue of the mammalian cancer-associated stem cell regulator Runx) as being an important regulator of seam development, acting to promote proliferation; rnt-1 mutants have fewer seam cells whereas overexpressing rnt-1 causes seam cell hyperplasia. We isolated the interacting CEH-20/Pbx and UNC-62/Meis TALE-class transcription factors during a genome-wide RNAi screen for novel regulators of seam cell number. Animals lacking wild type CEH-20 or UNC-62 display seam cell hyperplasia, largely restricted to the anterior of the worm, whereas double mutants have many additional seam cells along the length of the animal. The cellular basis of the hyperplasia involves the symmetrisation of normally asymmetric seam cell divisions towards the proliferative stem-like fate. The hyperplasia is completely suppressed in rnt-1 mutants, and rnt-1 is upregulated in ceh-20 and unc-62 mutants, suggesting that CEH-20 and UNC-62 function upstream of rnt-1 to limit proliferative potential to the appropriate daughter cell. In further support of this we find that CEH-20 is asymmetrically localised in seam daughters following an asymmetric division, being predominantly restricted to anterior nuclei whose fate is to differentiate. Thus, ceh-20 and unc-62 encode crucial regulators of seam cell division asymmetry, acting via rnt-1 to regulate the balance between proliferation and differentiation.
秀丽隐杆线虫的体壁 seam 细胞在幼虫发育过程中以类似于茎干的模式分裂,具有自我更新和产生分化后代的能力。 seam 细胞通常不对称分裂,产生一个与真皮融合的前体细胞和一个进一步增殖的后体细胞。以前,我们已经鉴定出 rnt-1(一种与哺乳动物癌症相关的干细胞调节因子 Runx 的同源物)作为 seam 发育的重要调节因子,促进增殖;rnt-1 突变体 seam 细胞较少,而过表达 rnt-1 会导致 seam 细胞增生。在一个针对 seam 细胞数量的新型调节因子的全基因组 RNAi 筛选中,我们分离了相互作用的 CEH-20/Pbx 和 UNC-62/Meis TALE 类转录因子。缺乏野生型 CEH-20 或 UNC-62 的动物表现出 seam 细胞增生,主要局限于线虫的前半部分,而双突变体在动物的全长上有许多额外的 seam 细胞。增生的细胞基础涉及到正常不对称 seam 细胞分裂向增殖性干细胞样命运的对称性。rnt-1 突变体完全抑制了增生,而 ceh-20 和 unc-62 突变体中 rnt-1 上调,这表明 CEH-20 和 UNC-62 在 rnt-1 上游发挥作用,将增殖潜力限制在适当的子细胞中。进一步支持这一点的是,我们发现 CEH-20 在不对称分裂后在 seam 后代中呈不对称定位,主要局限于将分化的前核。因此,ceh-20 和 unc-62 编码 seam 细胞分裂不对称的关键调节因子,通过 rnt-1 调节增殖和分化之间的平衡。