title: The Zic family homologue Odd-paired regulates Alk expression in Drosophila.
abstract: The Anaplastic Lymphoma Kinase (Alk) receptor tyrosine kinase (RTK) plays a critical role in the specification of founder cells (FCs) in the Drosophila visceral mesoderm (VM) during embryogenesis. Reporter gene and CRISPR/Cas9 deletion analysis reveals enhancer regions in and upstream of the Alk locus that influence tissue-specific expression in the amnioserosa (AS), the VM and the epidermis. By performing high throughput yeast one-hybrid screens (Y1H) with a library of Drosophila transcription factors (TFs) we identify Odd-paired (Opa), the Drosophila homologue of the vertebrate Zic family of TFs, as a novel regulator of embryonic Alk expression. Further characterization identifies evolutionarily conserved Opa-binding cis-regulatory motifs in one of the Alk associated enhancer elements. Employing Alk reporter lines as well as CRISPR/Cas9-mediated removal of regulatory elements in the Alk locus, we show modulation of Alk expression by Opa in the embryonic AS, epidermis and VM. In addition, we identify enhancer elements that integrate input from additional TFs, such as Binou (Bin) and Bagpipe (Bap), to regulate VM expression of Alk in a combinatorial manner. Taken together, our data show that the Opa zinc finger TF is a novel regulator of embryonic Alk expression.
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.|Department of Molecular Biology, Umeå University, Umeå, Sweden.
authors.forename: Patricia
authors.identifier: 0000-0002-6084-7962
authors.initials: P
authors.lastname: Mendoza-García
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Fredrik
authors.identifier: 0000-0002-8332-7011
authors.initials: F
authors.lastname: Hugosson
authors.affiliation: Department of Molecular Biology, Umeå University, Umeå, Sweden.
authors.forename: Mahsa
authors.initials: M
authors.lastname: Fallah
authors.affiliation: Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Higgins
authors.affiliation: Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: Yasuno
authors.initials: Y
authors.lastname: Iwasaki
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Pfeifer
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Georg
authors.initials: G
authors.lastname: Wolfstetter
authors.affiliation: Department of Molecular Biology, Umeå University, Umeå, Sweden.
authors.forename: Gaurav
authors.identifier: 0000-0002-0429-1904
authors.initials: G
authors.lastname: Varshney
authors.affiliation: Department of Molecular Biology, Umeå University, Umeå, Sweden.
authors.forename: Dmitry
authors.identifier: 0000-0001-8323-1717
authors.initials: D
authors.lastname: Popichenko
authors.affiliation: Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, United States of America.
authors.forename: J Peter
authors.identifier: 0000-0002-2122-2628
authors.initials: JP
authors.lastname: Gergen
authors.affiliation: Centre for Neural Circuits and Behaviour, University of Oxford, Oxford, United Kingdom.
authors.forename: Korneel
authors.identifier: 0000-0002-0362-7007
authors.initials: K
authors.lastname: Hens
authors.affiliation: Laboratory of Systems Biology and Genetics, Lausanne, Switzerland.
authors.forename: Bart
authors.identifier: 0000-0001-9935-843X
authors.initials: B
authors.lastname: Deplancke
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Ruth H
authors.identifier: 0000-0002-2735-8470
authors.initials: RH
authors.lastname: Palmer
mesh_terms: D033961:Adaptor Protein Complex 1 / Q000235:genetics / Q000378:metabolism; D033966:Adaptor Protein Complex beta Subunits / Q000235:genetics / Q000378:metabolism; D000077548:Anaplastic Lymphoma Kinase; D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites; D064113:CRISPR-Cas Systems; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology* / Q000235:genetics; D004625:Embryo, Nonmammalian; D004742:Enhancer Elements, Genetic; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D033961:Adaptor Protein Complex 1; D033966:Adaptor Protein Complex beta Subunits; C439125:Bap protein, Drosophila; D029721:Drosophila Proteins; D051858:Forkhead Transcription Factors; D018398:Homeodomain Proteins; C085443:Opa protein, Drosophila; D014157:Transcription Factors; C440017:bin protein, Drosophila; D000077548:Anaplastic Lymphoma Kinase; D020794:Receptor Protein-Tyrosine Kinases