Differentiation Driven Changes in the Dynamic Organization of Basal Transcription Initiation

Autor: Alex Maas, Catherine Miquel, Jan H.J. Hoeijmakers, Wim Vermeulen, Jan de Wit, Lise O. Andrieux, Nils Wijgers, Maria Fousteri, Pierre-Olivier Mari, Julie Nonnekens, Sophie Mourgues, Arjan F. Theil, Giuseppina Giglia-Mari
Přispěvatelé: Institut de pharmacologie et de biologie structurale (IPBS), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Department of Molecular Genetics [Rotterdam, The Netherlands] (Erasmus MC), Oncode Institute [Rotterdam, The Netherlands]-Cancer Genomics Netherlands [Rotterdam, The Netherlands], Service de Neuropathologie [Sainte-Anne], Hôpital Sainte-Anne, Erasmus University Medical Center [Rotterdam] (Erasmus MC), Université Paris 1 Panthéon-Sorbonne - UFR d'Arts plastiques et sciences de l'art (UP1 UFR04), Université Paris 1 Panthéon-Sorbonne (UP1), Department of Cell Biology and Genetics, National Cancer Institute [Bethesda] (NCI-NIH), National Institutes of Health [Bethesda] (NIH), Molecular Genetics, Cell biology
Rok vydání: 2009
Předmět:
Chromatin Immunoprecipitation
Transcription
Genetic
QH301-705.5
Recombinant Fusion Proteins
Cellular differentiation
Biology
General Biochemistry
Genetics and Molecular Biology
Mice
03 medical and health sciences
Organ Culture Techniques
0302 clinical medicine
Bacterial Proteins
Transcription (biology)
Cerebellum
Animals
Biology (General)
Transcription factor
Cells
Cultured
Embryonic Stem Cells
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
Cerebral Cortex
Genetics
0303 health sciences
Molecular Biology/DNA Repair
General Immunology and Microbiology
General transcription factor
General Neuroscience
fungi
food and beverages
Gene Expression Regulation
Developmental
Fluorescence recovery after photobleaching
Cell Differentiation
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Molecular biology
Cell Biology
Molecular Biology/Transcription Initiation and Activation
Fibroblasts
Cell biology
Chromatin
Mice
Inbred C57BL
Kinetics
Luminescent Proteins
Transcription Factor TFIIH
Transcription factor II H
General Agricultural and Biological Sciences
030217 neurology & neurosurgery
Research Article
Fluorescence Recovery After Photobleaching
Zdroj: PLoS Biology
PLoS Biology, Public Library of Science, 2009, 7 (10), pp.e1000220. ⟨10.1371/journal.pbio.1000220⟩
PLoS Biology, Vol 7, Iss 10, p e1000220 (2009)
PLoS Biology (print), 7(10). Public Library of Science
ISSN: 1545-7885
1544-9173
DOI: 10.1371/journal.pbio.1000220
Popis: A novel mouse model reveals that the dynamic behavior of transcription factors can vary considerably between different cells of an organism.
Studies based on cell-free systems and on in vitro–cultured living cells support the concept that many cellular processes, such as transcription initiation, are highly dynamic: individual proteins stochastically bind to their substrates and disassemble after reaction completion. This dynamic nature allows quick adaptation of transcription to changing conditions. However, it is unknown to what extent this dynamic transcription organization holds for postmitotic cells embedded in mammalian tissue. To allow analysis of transcription initiation dynamics directly into living mammalian tissues, we created a knock-in mouse model expressing fluorescently tagged TFIIH. Surprisingly and in contrast to what has been observed in cultured and proliferating cells, postmitotic murine cells embedded in their tissue exhibit a strong and long-lasting transcription-dependent immobilization of TFIIH. This immobilization is both differentiation driven and development dependent. Furthermore, although very statically bound, TFIIH can be remobilized to respond to new transcriptional needs. This divergent spatiotemporal transcriptional organization in different cells of the soma revisits the generally accepted highly dynamic concept of the kinetic framework of transcription and shows how basic processes, such as transcription, can be organized in a fundamentally different fashion in intact organisms as previously deduced from in vitro studies.
Author Summary The accepted model of eukaryotic mRNA production is that transcription factors spend most of their time diffusing throughout the cell nucleus, encountering gene promoters (their substrate) in a random fashion and binding to them for a very short time. A similar modus operandi has been accepted as a paradigm for interactions within most of the chromatin-associated enzymatic processes (transcription, replication, DNA damage response). However, it is not known whether such behavior is indeed a common characteristic for all cells in the organism. To answer this question, we generated a knock-in mouse that expresses in all cells a fluorescently tagged transcription factor (TFIIH) that functions in both transcription initiation and DNA repair. This new tool, when combined with quantitative imaging techniques, allowed us to monitor the mobility of this transcription factor in virtually all living tissues. In this study, we show that, in contrast to the aforementioned paradigm, in highly differentiated postmitotic cells such as neurons, hepatocytes, and cardiac myocytes, TFIIH is effectively immobilized on the chromatin during transcription, whereas in proliferative cells, TFIIH has the same dynamic behavior as in cultured cells. Our study also points out that results obtained from in vitro or cultured cell systems cannot always be directly extrapolated to the whole organism. More importantly, this raises a question for researchers in the transcription field: why do some cells opt for a dynamic framework for transcription, whereas others exhibit a static one?
Databáze: OpenAIRE
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