McGill Melanie A, McKinley R F Andrew, Harris Tony J C
Department of Cell and Systems Biology, University of Toronto, Toronto, Ontario M5S3G5, Canada.
J Cell Biol. 2009 Jun 1;185(5):787-96. doi: 10.1083/jcb.200812146. Epub 2009 May 25.
Proper epithelial structure requires adherens junction (AJ) assembly. In the early Drosophila embryo, AJ assembly depends on Bazooka (Baz; PAR-3), but it is unclear how Baz affects AJ assembly and what precursors are involved. To understand this process at the molecular level, we counted the number of core AJ proteins and Baz proteins at an average spot AJ (SAJ) and determined their dynamics with fluorescence recovery after photobleaching experiments. These data reveal that SAJs are subdivided into Baz clusters and cadherin-catenin clusters with independent protein numbers and dynamics. This independence suggests that precursory cadherin-catenin clusters might form before SAJ assembly. We identify cadherin-catenin clusters forming between apical microvilli. Further analyses show that they form independently of Baz and that Baz functions in repositioning them to apicolateral sites for full SAJ assembly. Our data implicate cell protrusions in initial cadherin-catenin clustering in the Drosophila melanogaster embryo. Then, independent Baz clusters appear to engage the cadherin-catenin clusters to assemble SAJs.
合适的上皮结构需要黏着连接(AJ)组装。在果蝇早期胚胎中,AJ组装依赖于巴左卡(Baz;PAR-3),但尚不清楚Baz如何影响AJ组装以及涉及哪些前体。为了在分子水平上理解这一过程,我们计算了平均斑点状AJ(SAJ)处核心AJ蛋白和Baz蛋白的数量,并通过光漂白后的荧光恢复实验确定了它们的动态变化。这些数据表明,SAJ可细分为具有独立蛋白数量和动态变化的Baz簇和钙黏蛋白-连环蛋白簇。这种独立性表明,前体钙黏蛋白-连环蛋白簇可能在SAJ组装之前形成。我们发现顶端微绒毛之间形成了钙黏蛋白-连环蛋白簇。进一步分析表明,它们独立于Baz形成,且Baz在将它们重新定位到顶侧位点以进行完整的SAJ组装中发挥作用。我们的数据表明,细胞突起参与了黑腹果蝇胚胎中最初的钙黏蛋白-连环蛋白簇集。然后,独立的Baz簇似乎与钙黏蛋白-连环蛋白簇结合以组装SAJ。