title: Inhibition of endogenous MTF-1 signaling in zebrafish embryos identifies novel roles for MTF-1 in development.
abstract: The metal responsive element-binding transcription factor-1 (MTF-1) responds to changes in cellular zinc levels caused by zinc exposure or disruption of endogenous zinc homeostasis by heavy metals or oxygen-related stress. Here we report the functional characterization of a complete zebrafish MTF-1 in comparison with the previously identified isoform lacking the highly conserved cysteine-rich motif (Cys-X-Cys-Cys-X-Cys) found in all other vertebrate MTF-1 orthologs. In an effort to develop novel molecular tools, a constitutively nuclear dominant-negative MTF-1 (dnMTF-1) was generated as tool for inhibiting endogenous MTF-1 signaling. The in vivo efficacy of the dnMTF-1 was determined by microinjecting in vitro transcribed dnMTF-1 mRNA into zebrafish embryos (1-2 cell stage) followed by transcriptomic profiling using an Agilent 4x44K array on 28- and 36-hpf embryos. A total of 594 and 560 probes were identified as differentially expressed at 28hpf and 36hpf, respectively, with interesting overlaps between timepoints. The main categories of genes affected by the inhibition of MTF-1 signaling were: nuclear receptors and genes involved in stress signaling, neurogenesis, muscle development and contraction, eye development, and metal homeostasis, including novel observations in iron and heme homeostasis. Finally, we investigate both the transcriptional activator and transcriptional repressor role of MTF-1 in potential novel target genes identified by transcriptomic profiling during early zebrafish development.
authors.affiliation: Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA.
authors.forename: Britton
authors.initials: B
authors.lastname: O'Shields
authors.affiliation: Andrew McArthur Consulting, Hamilton, Ontario L8S 3P6, Canada.
authors.forename: Andrew G
authors.initials: AG
authors.lastname: McArthur
authors.affiliation: Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Holowiecki
authors.affiliation: Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Kamper
authors.affiliation: Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Tapley
authors.affiliation: Department of Biological Sciences, University of Alabama, Tuscaloosa, AL 35487, USA. Electronic address: mjjenny@bama.ua.edu.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Jenny
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002104:Cadmium / Q000633:toxicity; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000378:metabolism*; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D063990:Gene Ontology; D006466:Hemopexin / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*; D000097809:Transcription Factor MTF-1
chemical_list: D002104:Cadmium; D004268:DNA-Binding Proteins; D006466:Hemopexin; D012333:RNA, Messenger; D014157:Transcription Factors; D000097809:Transcription Factor MTF-1
keywords: Embryonic development; MTF-1; Metal; Zebrafish; Zinc homeostasis
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