Phosphorylation and ubiquitination of dynamin-related proteins (AtDRP3A/3B) synergically regulate mitochondrial proliferation during mitosis.
Phosphorylation and ubiquitination of dynamin-related proteins (AtDRP3A/3B) synergically regulate mitochondrial proliferation during mitosis.
复制标题
动力相关蛋白 (AtDRP3A/3B) 的磷酸化和泛素化协同调节有丝分裂过程中的线粒体增殖。
DOI：
10.1111/j.1365-313x.2012.05052.x
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发表时间：
2012-10
期刊：
影响因子：
7.2
通讯作者：
Lin, Jinxing
中科院分区：
文献类型：
--
作者：
Wang, Feng;Liu, Peng;Zhang, Quan;Zhu, Jian;Chen, Tong;Arimura, Shin-ichi;Tsutsumi, Nobuhiro;Lin, Jinxing
在有丝分裂过程中，线粒体分裂和融合之间的平衡被破坏，但植物细胞中控制这一现象的机制仍然是谜。我们利用线粒体基质定位的Kaede蛋白（mt-Kaede）分析了BY-2悬浮细胞线粒体分裂的动力学。对mt-Kaede的光激活荧光分析表明，在有丝分裂过程中，裂变过程占主导地位。这一发现得到了by -2悬浮细胞在不同阶段线粒体大小分布的电镜分析的证实。与myc标记的动力蛋白相关蛋白3A/3B相互作用的细胞蛋白（AtDRP3A和AtDRP3B）用抗myc抗体偶联珠免疫沉淀，随后用微毛细管液相色谱-四极杆飞行时间质谱（CapLC Q-TOF） MS/MS鉴定。鉴定的蛋白质与细胞骨架（微管）、磷酸化或泛素化功能广泛相关。用CdkB/cyclin B抑制剂或丝氨酸/苏氨酸蛋白激酶抑制剂处理细胞可抑制AtDRP3A/AtDRP3B的有丝分裂磷酸化和中期线粒体分裂。通过dendra2标记的AtDRP3A/3B在绿到红光转换后的荧光延时成像来跟踪AtDRP3A/3B在细胞周期中的命运；实验表明AtDRP3A/3B在间期被部分降解。此外，我们发现微管在有丝分裂过程中参与线粒体分裂，线粒体向子细胞的运动早在中期就受到限制。综上所述，这些研究结果表明，AtDRP3A/3B的有丝分裂磷酸化促进了植物细胞有丝分裂过程中的线粒体分裂，并且AtDRP3A/3B在间期部分降解，从而为BY-2悬浮细胞中与细胞周期转变相关的线粒体形态变化提供了机制见解。
The balance between mitochondrial fission and fusion is disrupted during mitosis, but the mechanism governing this phenomenon in plant cells remains enigmatic. Here, we used mitochondrial matrix-localized Kaede protein (mt-Kaede) to analyze the dynamics of mitochondrial fission in BY-2 suspension cells. Analysis of the photoactivatable fluorescence of mt-Kaede suggested that the fission process is dominant during mitosis. This finding was confirmed by an electron microscopic analysis of the size distribution of mitochondria in BY-2 suspension cells at various stages. Cellular proteins interacting with Myc-tagged dynamin-related protein 3A/3B (AtDRP3A and AtDRP3B) were immunoprecipitated with anti-Myc antibody-conjugated beads and subsequently identified by microcapillary liquid chromatography-quadrupole time-of-flight mass spectrometry (CapLC Q-TOF) MS/MS. The identified proteins were broadly associated with cytoskeletal (microtubular), phosphorylation, or ubiquitination functions. Mitotic phosphorylation of AtDRP3A/AtDRP3B and mitochondrial fission at metaphase were inhibited by treatment of the cells with a CdkB/cyclin B inhibitor or a serine/threonine protein kinase inhibitor. The fate of AtDRP3A/3B during the cell cycle was followed by time-lapse imaging of the fluorescence of Dendra2-tagged AtDRP3A/3B after green-to-red photoconversion; this experiment showed that AtDRP3A/3B is partially degraded during interphase. Additionally, we found that microtubules are involved in mitochondrial fission during mitosis, and that mitochondria movement to daughter cell was limited as early as metaphase. Taken together, these findings suggest that mitotic phosphorylation of AtDRP3A/3B promotes mitochondrial fission during plant cell mitosis, and that AtDRP3A/3B is partially degraded at interphase, providing mechanistic insight into the mitochondrial morphological changes associated with cell-cycle transitions in BY-2 suspension cells.
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影响因子：
--
作者：
Jourdain, Isabelle;Gachet, Yannick;Hyams, Jeremy S.
通讯作者：
Hyams, Jeremy S.
影响因子：
4
作者：
Braschi, Emelie;McBride, Heidi M.
通讯作者：
McBride, Heidi M.
影响因子：
3.8
作者：
Gerbin, Candice S.;Landgraf, Ralf
通讯作者：
Landgraf, Ralf
影响因子：
7.4
作者：
Nebenführ, A;Frohlick, JA;Staehelin, LA
通讯作者：
Staehelin, LA
影响因子：
64.8
作者：
OBAR, RA;COLLINS, CA;VALLEE, RB
通讯作者：
VALLEE, RB