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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
Abstract. Objectives: This article is to study the role of G(1)/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 endoderm...
Autores principales: | , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Blackwell Publishing Ltd
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2659368/ https://www.ncbi.nlm.nih.gov/pubmed/19040567 http://dx.doi.org/10.1111/j.1365-2184.2008.00556.x |
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author | Bryja, V. Pacherník, J. Vondráček, J. Souček, K. Čajánek, L. Horvath, V. Holubcová, Z. Dvořák, P. Hampl, A. |
author_facet | Bryja, V. Pacherník, J. Vondráček, J. Souček, K. Čajánek, L. Horvath, V. Holubcová, Z. Dvořák, P. Hampl, A. |
author_sort | Bryja, V. |
collection | PubMed |
description | Abstract. Objectives: This article is to study the role of G(1)/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 G(1)/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 G(1)/S transition‐regulating machinery in early embryonic cells. |
format | Text |
id | pubmed-2659368 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Blackwell Publishing Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-26593682009-03-30 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 Bryja, V. Pacherník, J. Vondráček, J. Souček, K. Čajánek, L. Horvath, V. Holubcová, Z. Dvořák, P. Hampl, A. Cell Prolif Original Articles Abstract. Objectives: This article is to study the role of G(1)/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 G(1)/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 G(1)/S transition‐regulating machinery in early embryonic cells. Blackwell Publishing Ltd 2008-10-06 /pmc/articles/PMC2659368/ /pubmed/19040567 http://dx.doi.org/10.1111/j.1365-2184.2008.00556.x Text en © 2008 The Authors. Journal compilation © 2008 Blackwell Publishing Ltd Open access. |
spellingShingle | Original Articles Bryja, V. Pacherník, J. Vondráček, J. Souček, K. Čajánek, L. Horvath, V. Holubcová, Z. Dvořák, P. Hampl, A. 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 |
title | 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 |
title_full | 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 |
title_fullStr | 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 |
title_full_unstemmed | 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 |
title_short | 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 |
title_sort | 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 |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2659368/ https://www.ncbi.nlm.nih.gov/pubmed/19040567 http://dx.doi.org/10.1111/j.1365-2184.2008.00556.x |
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