Abstract
Fasciculation and elongation zeta-1 (FEZ1) protein is involved in axon outgrowth and is highly expressed in the brain. It has multiple interaction partners, with functions varying from the regulation of neuronal development and intracellular transport mechanisms to transcription regulation. One of its interactors is retinoic acid receptor (RAR), which is activated by retinoic acid and controls many target genes and physiological process. Based on previous evidence suggesting a possible nuclear role for FEZ1, we wanted to deepen our understanding of this function by addressing the FEZ1–RAR interaction. We performed in vitro binding experiments and assessed the interface of interaction between both proteins. We found that FEZ1–RAR interacted with a similar magnitude as RAR to its responsive element DR5 and that the interaction occurred in the coiled-coil region of FEZ1 and in the ligand-binding domain of RAR. Furthermore, cellular experiments were performed in order to confirm the interaction and screen for induced target genes from an 86-gene panel. The analysis of gene expression showed that only in the presence of retinoic acid did FEZ1 induce hoxb4 gene expression. This finding is consistent with data from the literature showing the hoxb4 gene functionally involved in development and acute myeloid leukemia, as is FEZ1.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 4-14 |
| Number of pages | 11 |
| Journal | FEBS Open Bio |
| Volume | 8 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 2018 |
Bibliographical note
Funding Information:We thank São Paulo Research Foundation (FAPESP, Funda©cão de Amparo á Pesquisa do Estado de São Paulo) for funding this project by grants to JK (2012/ 18796-3) and fellowships to MBT (2012/00792-1; 2015/ 11059-1). We also thank the National Council for Scientific and Technological Development (CNPq; Conselho Nacional de Desenvolvimento Científico e Tecnológico) for financial support. We thank Brazilian Biosciences National Laboratory and its facilities (Spectroscopy and Calorimetry, Mass Spectrometry (2009/54067-3), Protein Purification, Confocal Microscopy). We thank the University of Minnesota Medical School and its Genomic Center facility. We thank Dr. Bomi Lee for fruitful discussions and protocols sharing that certainly improved our work. We thank Dr. Edmarcia Elisa de Souza for sharing her knowledge about immunofluorescence. We also like to thank the National Institute of Science and Technology on Photonics Applied to Cell Biology (INFABIC, UNICAMP) for imaging support. INFABIC is cofunded by Funda©cão de Amparo a Pes-quisa do Estado de São Paulo (FAPESP, 08/57906-3) and National Council for Scientific and Technological Development (CNPq, 573913/2008-0).
Publisher Copyright:
© 2017 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.
Keywords
- FEZ
- RAR
- gene regulation
- nuclear function
- retinoic acid
- transcription
