PMID- 31690123 OWN - NLM STAT- MEDLINE DCOM- 20200615 LR - 20231213 IS - 1530-6860 (Electronic) IS - 0892-6638 (Print) IS - 0892-6638 (Linking) VI - 33 IP - 12 DP - 2019 Dec TI - The classic metal-sensing transcription factor MTF1 promotes myogenesis in response to copper. PG - 14556-14574 LID - 10.1096/fj.201901606R [doi] AB - Metal-regulatory transcription factor 1 (MTF1) is a conserved metal-binding transcription factor in eukaryotes that binds to conserved DNA sequence motifs, termed metal response elements. MTF1 responds to both metal excess and deprivation, protects cells from oxidative and hypoxic stresses, and is required for embryonic development in vertebrates. To examine the role for MTF1 in cell differentiation, we use multiple experimental strategies [including gene knockdown (KD) mediated by small hairpin RNA and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9), immunofluorescence, chromatin immunopreciptation sequencing, subcellular fractionation, and atomic absorbance spectroscopy] and report a previously unappreciated role for MTF1 and copper (Cu) in cell differentiation. Upon initiation of myogenesis from primary myoblasts, both MTF1 expression and nuclear localization increased. Mtf1 KD impaired differentiation, whereas addition of nontoxic concentrations of Cu(+)-enhanced MTF1 expression and promoted myogenesis. Furthermore, we observed that Cu(+) binds stoichiometrically to a C terminus tetra-cysteine of MTF1. MTF1 bound to chromatin at the promoter regions of myogenic genes, and Cu addition stimulated this binding. Of note, MTF1 formed a complex with myogenic differentiation (MYOD)1, the master transcriptional regulator of the myogenic lineage, at myogenic promoters. These findings uncover unexpected mechanisms by which Cu and MTF1 regulate gene expression during myoblast differentiation.-Tavera-Montanez, C., Hainer, S. J., Cangussu, D., Gordon, S. J. V., Xiao, Y., Reyes-Gutierrez, P., Imbalzano, A. N., Navea, J. G., Fazzio, T. G., Padilla-Benavides, T. The classic metal-sensing transcription factor MTF1 promotes myogenesis in response to copper. FAU - Tavera-Montanez, Cristina AU - Tavera-Montanez C AD - Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. FAU - Hainer, Sarah J AU - Hainer SJ AD - Department of Molecular, Cell, and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. FAU - Cangussu, Daniella AU - Cangussu D AD - Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. FAU - Gordon, Shellaina J V AU - Gordon SJV AD - Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. FAU - Xiao, Yao AU - Xiao Y AD - Department of Chemistry, Skidmore College, Saratoga Springs, New York, USA. FAU - Reyes-Gutierrez, Pablo AU - Reyes-Gutierrez P AD - Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. FAU - Imbalzano, Anthony N AU - Imbalzano AN AD - Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. FAU - Navea, Juan G AU - Navea JG AD - Department of Chemistry, Skidmore College, Saratoga Springs, New York, USA. FAU - Fazzio, Thomas G AU - Fazzio TG AD - Department of Molecular, Cell, and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. FAU - Padilla-Benavides, Teresita AU - Padilla-Benavides T AD - Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA. LA - eng GR - R01 GM056244/GM/NIGMS NIH HHS/United States GR - R01 HD072122/HD/NICHD NIH HHS/United States GR - R25 HL092610/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20191105 PL - United States TA - FASEB J JT - FASEB journal : official publication of the Federation of American Societies for Experimental Biology JID - 8804484 RN - 0 (DNA-Binding Proteins) RN - 0 (MyoD Protein) RN - 0 (MyoD1 myogenic differentiation protein) RN - 0 (Transcription Factors) RN - 789U1901C5 (Copper) SB - IM MH - Animals MH - *Cell Differentiation MH - Cells, Cultured MH - Copper/*pharmacology MH - DNA-Binding Proteins/*metabolism MH - Mice MH - Mice, Inbred C57BL MH - *Muscle Development MH - MyoD Protein/metabolism MH - Myoblasts/cytology/drug effects/*metabolism MH - Transcription Factors/*metabolism MH - Transcription Factor MTF-1 PMC - PMC6894080 OTO - NOTNLM OT - ChIP-Seq OT - copper binding OT - myogenic differentiation 1 COIS- The authors thank Courtney McCann, Dr. Sabriya Syed, Dr. Hanna Witwicka, and members of the T.P.-B. laboratory for insightful discussions regarding this manuscript. This work was supported by the Faculty Diversity Scholars Award from the University of Massachusetts Medical School to T.P.-B. Funding from the U.S. National Institute of Health [NIH; Grants R01GM56244 (National Institute of General Medical Sciences) to A.N.I. and R01HD072122 (Eunice Kennedy Shriver National Institute of Child Health and Human Development) to T.G.F.). S.J.V.G was supported by the University of Massachusetts Medical School funding for the Summer Undergraduate Research Experience Program [NIH Grant 2R25HL092610 (National Heart, Lung, and Blood Institute)]. Funding from the National Science Foundation [Division of Biological Infrastructure (DBI) 0959476 to J.G.N.]. S.J.H. is a Special Fellow of the Leukemia and Lymphoma Society. The authors declare no conflicts of interest. EDAT- 2019/11/07 06:00 MHDA- 2020/06/17 06:00 PMCR- 2020/11/05 CRDT- 2019/11/07 06:00 PHST- 2019/11/07 06:00 [pubmed] PHST- 2020/06/17 06:00 [medline] PHST- 2019/11/07 06:00 [entrez] PHST- 2020/11/05 00:00 [pmc-release] AID - FJ_201901606R [pii] AID - 10.1096/fj.201901606R [doi] PST - ppublish SO - FASEB J. 2019 Dec;33(12):14556-14574. doi: 10.1096/fj.201901606R. Epub 2019 Nov 5.