An evolutionarily conserved kernel of gata5, gata6, otx2 and prdm1a operates in the formation of endoderm in zebrafish

Autor: Chiou-Hwa Yuh, Chang-Ben Huang, Te-Hsuan Jang, Wen-Fang Tseng
Rok vydání: 2011
Předmět:
endocrine system
animal structures
Time Factors
Morpholino
GATA5 Transcription Factor
DNA Mutational Analysis
Molecular Sequence Data
Gene regulatory network
Biology
GATA Transcription Factors
Conserved sequence
Evolution
Molecular
medicine
Animals
Gene Regulatory Networks
RNA
Messenger
Zebrafish
Molecular Biology
Conserved Sequence
Body Patterning
Genetics
Otx Transcription Factors
Base Sequence
Gene Expression Profiling
Endoderm
Gene Expression Regulation
Developmental
Nuclear Proteins
Reproducibility of Results
Mesendoderm
Epistasis
Genetic
Cell Biology
Oligonucleotides
Antisense
Zebrafish Proteins
biology.organism_classification
Cell biology
Gene expression profiling
DNA-Binding Proteins
Kernel
medicine.anatomical_structure
Genetic Loci
embryonic structures
GATA transcription factor
Positive Regulatory Domain I-Binding Factor 1
Chromatin immunoprecipitation
Protein Binding
Developmental Biology
Zdroj: Developmental Biology. 357(2):541-557
ISSN: 0012-1606
DOI: 10.1016/j.ydbio.2011.06.040
Popis: An evolutionarily conserved subcircuit (kernel) dedicated to a specific developmental function is found at the top of the gene regulatory networks (GRNs) hierarchy. Here we comprehensively demonstrate that a pan-deuterostome endoderm specification kernel exists in zebrafish. We analyzed interactions among gata5, gata6, otx2 and prdm1a using specific morpholino knockdowns and measured the gene expression profiles by quantitative real-time RT-PCR and in situ hybridization. The mRNA rescue experiment validated the specificity of the morpholino knockdown. We found that the interactions among gata5, gata6, otx2 and prdm1a determine the initial specification of the zebrafish endoderm. Although otx2 can activate both gata5 and gata6, and the prdm1a/krox homologue also activates some endoderm transcription factors, a feedback loop from Gata to otx2 and prdm1a is missing. Furthermore, we found the positive regulation between gata5 and gata6 to further lock-on the mesendoderm specification by the Gata family. Chromatin immunoprecipitation was used to further validate the recruitment of Otx2 to the gata5 and gata6 loci. Functional assays revealed that module B of gata6 and the basal promoter of gata5 drive the gene at the mesendoderm, and mutational analysis demonstrated that Otx2 and Gata5/6 contribute to reporter gene activation. This is the first direct evidence for evolutionarily conserved endoderm specification across echinoderms and vertebrates.
Databáze: OpenAIRE
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