Modulation of the Fanconi anemia pathway via chemically induced changes in chromatin structure.

David A Vierra,Maurizio Mauro,Morganne M Adroved,Meghan A Rego,Niall G Howlett,Jada L Garzon

doi:10.18632/oncotarget.19470

David A Vierra, Maurizio Mauro + Show 4 more

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https://doi.org/10.18632/oncotarget.19470

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Abstract

Fanconi anemia (FA) is a rare disease characterized by congenital defects, bone marrow failure, and atypically early-onset cancers. The FA proteins function cooperatively to repair DNA interstrand crosslinks. A major step in the activation of the pathway is the monoubiquitination of the FANCD2 and FANCI proteins, and their recruitment to chromatin-associated nuclear foci. The regulation and function of FANCD2 and FANCI, however, is poorly understood. In addition, how chromatin state impacts pathway activation is also unknown. In this study, we have examined the influence of chromatin state on the activation of the FA pathway. We describe potent activation of FANCD2 and FANCI monoubiquitination and nuclear foci formation following treatment of cells with the histone methyltransferase inhibitor BRD4770. BRD4770-induced activation of the pathway does not occur via the direct induction of DNA damage or via the inhibition of the G9a histone methyltransferase, a mechanism previously proposed for this molecule. Instead, we show that BRD4770-inducible FANCD2 and FANCI monoubiquitination and nuclear foci formation may be a consequence of inhibition of the PRC2/EZH2 chromatin-modifying complex. In addition, we show that inhibition of the class I and II histone deacetylases leads to attenuated FANCD2 and FANCI monoubiquitination and nuclear foci formation. Our studies establish that chromatin state is a major determinant of the activation of the FA pathway and suggest an important role for the PRC2/EZH2 complex in the regulation of this critical tumor suppressor pathway.

Highlights

All organisms are continuously exposed to endogenous and exogenous DNA damaging agents, including reactive oxygen species and aldehydes from normal metabolic processes and UV irradiation from sunlight
We show that BRD4770inducible FANCD2 and FANCI monoubiquitination and nuclear foci formation may be a consequence of inhibition of the PRC2/EZH2 chromatin-modifying complex
Inhibition of class I and II histone deacetylase (HDAC) attenuates activation of the Fanconi anemia (FA) pathway To investigate the influence of histone acetylation state on the activation of the FA pathway, we examined the effects of the class I and II HDAC inhibitors trichostatin A (TSA) and vorinostat (SAHA) on FANCD2 and FANCI monoubiquitination and nuclear foci formation

Summary

Introduction

All organisms are continuously exposed to endogenous and exogenous DNA damaging agents, including reactive oxygen species and aldehydes from normal metabolic processes and UV irradiation from sunlight. The timely and accurate repair of DNA damage is essential for the maintenance of genome stability and organismal survival. Chromatin represents the higher order macromolecular complex of DNA and histone proteins, and chromatin plasticity has become increasingly recognized as a major determinant of DNA damage recognition, signaling, and repair [1,2,3]. The nucleosome is the fundamental subunit of chromatin and exhibits plasticity via compositional alteration, translational repositioning, and the posttranslational modification of histone tails. Histone tails are subject www.impactjournals.com/oncotarget to a wide variety of posttranslational modifications including acetylation, methylation, phosphorylation, and ubiquitination [4]. Underscoring the importance of histone acetylation in DNA repair, key roles for TIP60/KAT5, HDAC1, and HDAC2 in the maintenance of genome stability have been established [5,6,7]

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