Akap350 Recruits Eb1 to The Spindle Poles, Ensuring Proper Spindle Orientation and Lumen Formation in 3d Epithelial Cell Cultures.
Akap350 Recruits Eb1 to The Spindle Poles, Ensuring Proper Spindle Orientation and Lumen Formation in 3d Epithelial Cell Cultures.
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DOI：
10.1038/s41598-017-14241-y
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发表时间：
2017-11-02
影响因子：
4.6
通讯作者：
Larocca MC
中科院分区：
文献类型：
--
作者：
Almada E;Tonucci FM;Hidalgo F;Ferretti A;Ibarra S;Pariani A;Vena R;Favre C;Girardini J;Kierbel A;Larocca MC
上皮细胞的组织形成具有单腔的中空器官需要细胞分裂的精确三维排列。有丝分裂纺锤体的方向是由信号通路来定义的，这些信号通路提供了细胞皮层和星形微管上特定位点之间的分子联系，这一点尚未完全阐明。AKAP 350是一种中心体/高尔基体支架蛋白，参与微管动力学的调节。使用3D上皮细胞培养物，我们发现AKAP 350表达降低的细胞（AKAP 350 KD）形成具有异常管腔形态的极化囊肿。AKAP 350 KD包囊中有丝分裂细胞的分析表明纺锤体排列有缺陷。我们确定AKAP 350与EB 1相互作用，EB 1是一种微管相关蛋白，在纺锤体极点调节纺锤体方向。AKAP 350表达的减少导致纺锤体极和星形微管的EB 1水平显著降低。相反，EB 1的过表达挽救了由AKAP 350表达缺陷诱导的纺锤体方向缺陷。AKAP 350/EB 1复合物从中心体的特异性离域降低了星形微管的EB 1水平，并导致类似AKAP 350 KD包囊的3D器官型结构的形成。我们的结论是，AKAP 350招聘EB 1的纺锤体极点，确保EB 1的存在在星形微管和适当的纺锤体方向在上皮形态发生。
The organization of epithelial cells to form hollow organs with a single lumen requires the accurate three-dimensional arrangement of cell divisions. Mitotic spindle orientation is defined by signaling pathways that provide molecular links between specific spots at the cell cortex and astral microtubules, which have not been fully elucidated. AKAP350 is a centrosomal/Golgi scaffold protein, implicated in the regulation of microtubule dynamics. Using 3D epithelial cell cultures, we found that cells with decreased AKAP350 expression (AKAP350KD) formed polarized cysts with abnormal lumen morphology. Analysis of mitotic cells in AKAP350KD cysts indicated defective spindle alignment. We established that AKAP350 interacts with EB1, a microtubule associated protein that regulates spindle orientation, at the spindle poles. Decrease of AKAP350 expression lead to a significant reduction of EB1 levels at spindle poles and astral microtubules. Conversely, overexpression of EB1 rescued the defective spindle orientation induced by deficient AKAP350 expression. The specific delocalization of the AKAP350/EB1complex from the centrosome decreased EB1 levels at astral microtubules and lead to the formation of 3D-organotypic structures which resembled AKAP350KD cysts. We conclude that AKAP350 recruits EB1 to the spindle poles, ensuring EB1 presence at astral microtubules and proper spindle orientation during epithelial morphogenesis.
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影响因子：
5.6
作者：
Mattaloni, Stella M.;Kolobova, Elena;Favre, Cristian;Marinelli, Raul A.;Goldenring, James R.;Larocca, Maria C.
通讯作者：
Larocca, Maria C.
影响因子：
9.2
作者：
Gierke, Sarah;Wittmann, Torsten
通讯作者：
Wittmann, Torsten
DOI：
10.1083/jcb.142.1.69
发表时间：
1998-07-13
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kenworthy AK;Edidin M
通讯作者：
Edidin M
影响因子：
21.3
作者：
通讯作者：
--
影响因子：
7.8
作者：
Martinez-Campos, Maruxa;Basto, Renata;Baker, James;Kernan, Maurice;Raff, Jordan W
通讯作者：
Raff, Jordan W