ZFIN ID: ZDB-PUB-030115-2
Pineal expression-promoting element (PIPE), a cis-acting element, directs pineal-specific gene expression in zebrafish
Asaoka, Y., Mano, H., Kojima, D., and Fukada, Y.
Date: 2002
Source: Proceedings of the National Academy of Sciences of the United States of America   99: 15456-15461 (Journal)
Registered Authors: Fukada, Yoshitaka, Kojima, Daisuke
Keywords: none
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified
  • Binding Sites
  • Eels/genetics
  • Eye Proteins/biosynthesis
  • Eye Proteins/genetics
  • Gene Expression Regulation/genetics*
  • Genes, Homeobox/genetics
  • Genes, Reporter
  • Green Fluorescent Proteins
  • Homeodomain Proteins/metabolism
  • Luminescent Proteins/biosynthesis
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Nerve Tissue Proteins/biosynthesis
  • Nerve Tissue Proteins/genetics
  • Organ Specificity
  • Otx Transcription Factors
  • Pineal Gland/metabolism*
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins/biosynthesis
  • Regulatory Sequences, Nucleic Acid*
  • Retina/metabolism
  • Rhodopsin/genetics
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Trans-Activators/metabolism
  • Zebrafish/genetics*
PubMed: 12438694 Full text @ Proc. Natl. Acad. Sci. USA
FIGURES
ABSTRACT
The pineal gland, sharing morphological and biochemical similarities with the retina, plays a unique and central role in the photoneuroendocrine system. The unique development of the pineal gland is directed by a specific combination of the expressed genes, but little is known about the regulatory mechanism underlying the pineal-specific gene expression. We isolated a 1.1-kbp fragment upstream of the zebrafish exo-rhodopsin (exorh) gene, which is expressed specifically in the pineal gland. Transgenic analysis using an enhanced green fluorescent protein reporter gene demonstrated that the proximal 147-bp region of the exorh promoter is sufficient to direct pineal-specific expression. This region contains three copies of a putative cone rod homeobox (Crx)Otx-binding site, which is known to be required for expression of both retina- and pineal-specific genes. Deletion and mutational analyses of the exorh promoter revealed that a previously uncharacterized sequence TGACCCCAATCT termed pineal expression-promoting element (PIPE) is required for pineal-specific promoter activity in addition to the CrxOtx-binding sites. By using the zebrafish rhodopsin (rh) promoter that drives retina-specific expression, we created a reporter construct having ectopic PIPE in the rh promoter at a position equivalent to that in the exorh promoter by introducing five nucleotide changes. Such a slight modification in the rh promoter induced ectopic enhanced green fluorescent protein expression in the pineal gland without affecting its retinal expression. These results identify PIPE as a critical cis-element contributing to the pineal-specific gene expression, in combination with the CrxOtx-binding site(s).
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