Lineage specific composition of cyclin D-CDK4/CDK6-p27 complexes reveals distinct functions of CDK4, CDK6 and individual D-type cyclins in differentiating cells of embryonic origin.
Lineage specific composition of cyclin D-CDK4/CDK6-p27 complexes reveals distinct functions of CDK4, CDK6 and individual D-type cyclins in differentiating cells of embryonic origin.
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DOI：
10.1111/j.1365-2184.2008.00556.x
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发表时间：
2008-12
影响因子：
8.5
通讯作者：
Hampl A
中科院分区：
文献类型：
--
作者：
Bryja V;Pacherník J;Vondráček J;Souček K;Čajánek L;Horvath V;Holubcová Z;Dvořák P;Hampl A
摘要：目的：研究G1/S期调控因子在多能性胚胎细胞分化中的作用。 材料与方法：我们建立了一个基于P19胚胎癌细胞的实验系统，该系统受益于两种类似的分化方案，产生内胚层或神经外胚层谱系。相对于细胞的生长和分化参数，分析了G1/S调节剂的水平、相互作用、活性和定位。结果与结论：我们证明，分化细胞的增殖参数与细胞周期蛋白A/E-CDK 2的活性和结构相关，但与细胞周期蛋白D-CDK 4/6-p27复合物无关。在指数生长的P19细胞群中，检测到细胞周期蛋白D1-CDK 4复合物，其在密度停滞后被细胞周期蛋白D2/3-CDK 4/6-p27复合物取代。在内胚层分化过程中，形成激酶失活的细胞周期蛋白D2/D3-CDK 4-p27复合物。神经分化特异性地诱导细胞周期蛋白D1，以细胞周期蛋白D3为代价，并导致细胞周期蛋白D1/D2-CDK 4-p27复合物的主要形成。分化伴随着细胞周期蛋白Ds和CDK 4/6的细胞质积累，其在神经细胞中与神经生长相关。这里发现的大多数现象都可以在小鼠胚胎干细胞中重现。总之，我们的数据表明：（i）单个细胞周期蛋白D亚型特异性地用于细胞谱系，（ii）CDK 4和CDK 6的功能存在根本差异，以及（iii）细胞周期蛋白D-CDK 4/6复合物在分化细胞的细胞质中起作用。我们的研究揭示了早期胚胎细胞中G1/S转换调节机制的另一个复杂性水平。
Abstract. Objectives: This article is to study the role of G1/S regulators in differentiation of pluripotent embryonic cells. Materials and methods: We established a P19 embryonal carcinoma cell‐based experimental system, which profits from two similar differentiation protocols producing endodermal or neuroectodermal lineages. The levels, mutual interactions, activities, and localization of G1/S regulators were analysed with respect to growth and differentiation parameters of the cells. Results and Conclusions: We demonstrate that proliferation parameters of differentiating cells correlate with the activity and structure of cyclin A/E–CDK2 but not of cyclin D–CDK4/6–p27 complexes. In an exponentially growing P19 cell population, the cyclin D1–CDK4 complex is detected, which is replaced by cyclin D2/3–CDK4/6–p27 complex following density arrest. During endodermal differentiation kinase‐inactive cyclin D2/D3–CDK4–p27 complexes are formed. Neural differentiation specifically induces cyclin D1 at the expense of cyclin D3 and results in predominant formation of cyclin D1/D2–CDK4–p27 complexes. Differentiation is accompanied by cytoplasmic accumulation of cyclin Ds and CDK4/6, which in neural cells are associated with neural outgrowths. Most phenomena found here can be reproduced in mouse embryonic stem cells. In summary, our data demonstrate (i) that individual cyclin D isoforms are utilized in cells lineage specifically, (ii) that fundamental difference in the function of CDK4 and CDK6 exists, and (iii) that cyclin D–CDK4/6 complexes function in the cytoplasm of differentiated cells. Our study unravels another level of complexity in G1/S transition‐regulating machinery in early embryonic cells.
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影响因子：
64.8
作者：
Andang, Michael;Hjerling-Leffler, Jens;Ernfors, Patrik
通讯作者：
Ernfors, Patrik
DOI：
10.1016/j.bbamcr.2004.01.001
发表时间：
2004-05-03
影响因子：
5.1
作者：
Bryja, V;Pacherník, J;Hampl, A
通讯作者：
Hampl, A
影响因子：
5.2
作者：
Bryja, V;Cajánek, L;Hampl, A
通讯作者：
Hampl, A
影响因子：
64.5
作者：
Kozar, K;Ciemerych, MA;Sicinski, P
通讯作者：
Sicinski, P
影响因子：
10.5
作者：
Ciemerych, MA;Kenney, AM;Sicinski, P
通讯作者：
Sicinski, P