Fol. Biol. 2018, 64, 1-9
Valproic Acid Decreases the Nuclear Localization of MDT-28, the Nematode Orthologue of MED28
Mediator is a multiprotein complex that connects regulation mediated by transcription factors with RNA polymerase II transcriptional machinery and integrates signals from the cell regulatory cascades with gene expression. One of the Mediator subunits, Mediator complex subunit 28 (MED28), has a dual nuclear and cytoplasmic localization and function. In the nucleus, MED28 functions as part of Mediator and in the cytoplasm, it interacts with cytoskeletal proteins and is part of the regulatory cascades including that of Grb2. MED28 thus has the potential to bring cytoplasmic regulatory interactions towards the centre of gene expression regulation. In this study, we identified MDT-28, the nematode orthologue of MED28, as a likely target of lysine acetylation using bioinformatic prediction of posttranslational modifications. Lysine acetylation was experimentally confirmed using anti-acetyl lysine antibody on immunoprecipitated GFP::MDT-28 expressed in synchronized C. elegans. Valproic acid (VPA), a known inhibitor of lysine deacetylases, enhanced the lysine acetylation of GFP::MDT-28. At the subcellular level, VPA decreased the nuclear localization of GFP::MDT-28 detected by fluorescencelifetime imaging microscopy (FLIM). This indicates that the nuclear pool of MDT-28 is regulated by a mechanism sensitive to VPA and provides an indirect support for a variable relative proportion of MED28 orthologues with other Mediator subunits.
Keywords
Mediator complex, valproic acid, nuclear localization, MDT-28, MED28.
Funding
This work was supported by the European Regional Development Fund “BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University in Vestec” (CZ.1.05/ 1.1.00/02.0109) and the LQ1604 National Sustainability Program II (Project BIOCEV-FAR) and the project Biocev (CZ.1.05/1.1.00/ 02.0109) from the Ministry of Education, Youth and Sports of the Czech Republic; PROGRES Q26/LF1 and PROGRES Q28 “Oncology” from Charles University in Prague; grant SVV 260377/2017 from Charles University in Prague. The imaging was done at the Imaging Methods Core Facility at Biocev, Faculty of Science, Charles University, supported by the Czech-BioImaging large RI project (LM2015062 funded by MEYS CR). AB acknowledges the support by the MEYS CR (CZ.02.1.01/ 0.0/0.0/16 013/0001775 Czech-Bioimaging).
References
Copyright
This is an open-access article distributed under the terms of the Creative Commons Attribution License.
