Briesewitz R, Kern A, Marcantonio E E
Department of Pathology, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Mol Biol Cell. 1995 Aug;6(8):997-1010. doi: 10.1091/mbc.6.8.997.
The membrane proximal regions of integrin alpha and beta subunits are highly conserved in evolution. In particular, all integrin alpha subunits share the KXGFFKR sequence at the beginning of their cytoplasmic domains. Previous work has shown that this domain is important in integrin receptor assembly. Using chimeric integrin alpha and beta subunits, we show that the native cytoplasmic domains of both subunits must be present for efficient assembly. Most strikingly, chimeric alpha 1 and beta 1 subunits with reciprocally swapped intracellular domains dimerize selectively into collagen IV receptors expressed at high levels on the surface. However, these receptors, which bind ligand efficiently, are deficient in a variety of post-ligand binding events, including cytoskeletal association and induction of tyrosine phosphorylation. Furthermore, deletion of the distal alpha cytoplasmic domain in the swapped heterodimers leads to ligand-independent focal contact localization, which also occurs in wild-type subunits when the distal cytoplasmic domain is deleted. These results show that proper integrin assembly requires opposed alpha and beta cytoplasmic domains, and this opposition prevents ligand-independent focal contact localization. Our working hypothesis is that these two domains may associate during receptor assembly and provide the mechanism for integrin receptor latency.
整合素α和β亚基的膜近端区域在进化过程中高度保守。特别是,所有整合素α亚基在其胞质结构域起始处都共享KXGFFKR序列。先前的研究表明,该结构域在整合素受体组装中很重要。利用嵌合整合素α和β亚基,我们发现两个亚基的天然胞质结构域都必须存在才能实现高效组装。最引人注目的是,细胞内结构域相互交换的嵌合α1和β1亚基选择性地二聚化形成在表面高水平表达的IV型胶原受体。然而,这些能有效结合配体的受体在多种配体结合后事件中存在缺陷,包括细胞骨架结合和酪氨酸磷酸化的诱导。此外,在交换的异二聚体中删除α亚基胞质结构域的远端会导致不依赖配体的粘着斑定位,当删除野生型亚基的远端胞质结构域时也会出现这种情况。这些结果表明,整合素的正确组装需要α和β胞质结构域相互对应,这种对应可防止不依赖配体的粘着斑定位。我们的工作假设是,这两个结构域可能在受体组装过程中相互作用,并为整合素受体的潜伏状态提供机制。