Chromatin plays a major role in the tight regulation of gene expression and in constraining inappropriate gene activity. Replication-coupled chromatin assembly ensures maintenance of these functions of chromatin during S phase of the cell cycle. Thus treatment of cells with an inhibitor of translation, such as cycloheximide (CX), would be expected to have a dramatic effect on chromatin structure and function, essentially in S phase of the cell cycle, due to uncoupled DNA replication and chromatin assembly. In this work, we confirm this hypothesis and show that CX can induce a dramatic S-phase-dependent alteration in chromatin structure that is associated with general RNA polymerase II-dependent transcriptional activation. Using two specific RNA polymerase II-transcribed genes, we confirm the above conclusion and show that CX-mediated transcriptional activation is enhanced during the DNA replication phase of the cell cycle. Moreover, we show co-operation between an inhibitor of histone deacetylase and CX in inducing gene expression, which is again S-phase-dependent. The modest effect of CX in inducing the activity of a transiently transfected promoter shows that the presence of the promoter in an endogenous chromatin context is necessary in order to observe transcriptional activation. We therefore suggest that the uncoupled DNA replication and histone synthesis that occur after CX treatment induces a general modification of chromatin structure, and propose that this general disorganization of chromatin structure is responsible for a widespread activation of RNA polymerase II-mediated gene transcription.
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December 1998
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Research Article|
December 15 1998
S-phase-dependent action of cycloheximide in relieving chromatin-mediated general transcriptional repression
Maya CESARI;
Maya CESARI
*Université de la Reunion, Laboratoire de Biochimie, 15 Av. René Cassin, 97489 St Denis Cedex, France
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Laurent HÉLIOT;
Laurent HÉLIOT
†Laboratoire de Biologie Moléculaire du Cycle Cellulaire–INSERM U309, Institut Albert Bonniot Faculté de Médecine, Domaine de la Merci, 38706 La Tronche Cedex, France
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Catherine MEPLAN;
Catherine MEPLAN
†Laboratoire de Biologie Moléculaire du Cycle Cellulaire–INSERM U309, Institut Albert Bonniot Faculté de Médecine, Domaine de la Merci, 38706 La Tronche Cedex, France
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Michel PABION;
Michel PABION
*Université de la Reunion, Laboratoire de Biochimie, 15 Av. René Cassin, 97489 St Denis Cedex, France
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Saadi KHOCHBIN
Saadi KHOCHBIN
1
†Laboratoire de Biologie Moléculaire du Cycle Cellulaire–INSERM U309, Institut Albert Bonniot Faculté de Médecine, Domaine de la Merci, 38706 La Tronche Cedex, France
1To whom correspondence should be addressed (e-mail [email protected]).
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Publisher: Portland Press Ltd
Received:
August 11 1998
Accepted:
October 02 1998
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 336 (3): 619–624.
Article history
Received:
August 11 1998
Accepted:
October 02 1998
Citation
Maya CESARI, Laurent HÉLIOT, Catherine MEPLAN, Michel PABION, Saadi KHOCHBIN; S-phase-dependent action of cycloheximide in relieving chromatin-mediated general transcriptional repression. Biochem J 15 December 1998; 336 (3): 619–624. doi: https://doi.org/10.1042/bj3360619
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