This proposal aims to study how the mitotic membrane network that surrounds the spindle microtubules regulates spindle morphogenesis and spindle orientation. The assembly and maintenance of mitotic spindle morphology requires balancing of forces generated by microtubule-based motor proteins such as dynein and the kinesin Eg5, and proper regulation of microtubule dynamics. Our studies of the lamin-B-containing mitotic membrane network, which we refer to as the lamin-B spindle sheath, have shown that this spindle-associated structure regulates spindle morphology and spindle orientation. Since lamin-B interacts with the dynein regulator NudEL and the microtubule depolymerase MCAK, we hypothesize that the lamin-B spindle sheath that surrounds the body of the spindle microtubules functions as a barrier to limit microtubule growth within the spindle boundary through MCAK and/or NudEL. Moreover, we propose that the lamin-B spindle sheath surrounding the spindle poles regulates the astral microtubules to ensure proper search and capture of the cortical spindle orientation cues. We will test these ideas in Aim 1 and Aim 2 using a number of assays and tools we have generated. We have shown previously that RanGTP, microtubules, and dynein are all required for the lamin-B spindle sheath assembly. More recently, we have found that lamin-B binds to several nucleoporins in mitosis.
In Aim 3, we will test the hypothesis that the interactions between lamin-B and nucleoporins are required for the assembly of the spindle sheath. These studies will make a significant contribution toward understanding how non-microtubule cellular structures regulate spindle assembly and orientation in mitosis.
Most studies of spindle assembly have focused on activities and forces within the spindle microtubules. However, many observations have shown that structures outside of spindle microtubules also contribute toward maintaining spindle morphology and position. Our studies have uncovered that a lamin-B-containing membrane network organized around the spindle microtubules plays a role in spindle morphogenesis. With the newly identified molecular handles, we propose to further dissect the mechanism by which the lamin- membrane network interacts with spindle microtubules to facilitate their organization and how this would in turn regulate spindle orientation during cell division. This proposal addresses an area of cell division that has received relatively little attention and it aims to uncover novel insights about spindle assembly mechanisms that had not been considered previously. Since the mitotic lamin-containing membrane network contains signaling molecules and cell fate determinants, we believe that understanding how this structure interacts with spindle microtubules during cell division will open new avenues to explore therapeutic targets for halting uncontrolled cell divisions.
|Chen, Haiyang; Zheng, Xiaobin; Zheng, Yixian (2014) Age-associated loss of lamin-B leads to systemic inflammation and gut hyperplasia. Cell 159:829-43|
|Jiang, Hao; He, Xiaonan; Wang, Shusheng et al. (2014) A microtubule-associated zinc finger protein, BuGZ, regulates mitotic chromosome alignment by ensuring Bub3 stability and kinetochore targeting. Dev Cell 28:268-81|
|Shi, Changji; Channels, Wilbur E; Zheng, Yixian et al. (2014) A computational model for the formation of lamin-B mitotic spindle envelope and matrix. Interface Focus 4:20130063|
|Liu, Zhonghua; Vong, Queenie P; Liu, Chengyu et al. (2014) Borg5 is required for angiogenesis by regulating persistent directional migration of the cardiac microvascular endothelial cells. Mol Biol Cell 25:841-51|
|Guo, Yuxuan; Kim, Youngjo; Shimi, Takeshi et al. (2014) Concentration-dependent lamin assembly and its roles in the localization of other nuclear proteins. Mol Biol Cell 25:1287-97|
|Kim, Youngjo; Zheng, Xiaobin; Zheng, Yixian (2013) Proliferation and differentiation of mouse embryonic stem cells lacking all lamins. Cell Res 23:1420-3|
|Zheng, Yixian; Iglesias, Pablo A (2013) Nucleating new branches from old. Cell 152:669-70|
|Kim, Youngjo; Zheng, Yixian (2013) Generation and characterization of a conditional deletion allele for Lmna in mice. Biochem Biophys Res Commun 440:8-13|
|Wang, Shusheng; Ketcham, Stephanie A; Schön, Arne et al. (2013) Nudel/NudE and Lis1 promote dynein and dynactin interaction in the context of spindle morphogenesis. Mol Biol Cell 24:3522-33|
|Chen, Haiyang; Chen, Xin; Zheng, Yixian (2013) The nuclear lamina regulates germline stem cell niche organization via modulation of EGFR signaling. Cell Stem Cell 13:73-86|
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