Ultraconserved non-coding sequence element controls a subset of spatiotemporal GLI3 expression.
The zinc-finger transcription factor GLI3 acts during vertebrate development in a combinatorial, context-dependent fashion as a primary transducer of sonic hedgehog (SHH) signaling. In humans, mutations affecting this key regulator of development are associated with GLI3-morphopathies, a group of congenital malformations in which forebrain and limb development are preferentially affected. We show that a non-coding element from intron two of GLI3, ultraconserved in mammals and highly conserved in the pufferfish Fugu, is a transcriptional enhancer. In transient transfection assays, it activates reporter gene transcription in human cell cultures expressing endogenous GLI3 but not in GLI3 negative cells. The identified enhancer element is predicted to contain conserved binding sites for transcription factors crucial for developmental steps in which GLI3 is involved. The regulatory potential of this element is conserved and was used to direct tissue-specific expression of a green fluorescent protein reporter gene in zebrafish embryos and of a beta-galactosidase reporter in transgenic mouse embryos. Time, location, and quantity of reporter gene expression are congruent with part of the pattern previously reported for endogenous GLI3 transcription.
Institute of Human Genetics, Philipps-University, Bahnhofstrasse 7, D35037 Marburg, Germany.
Zissis
Z
Paparidis
Amir Ali
AA
Abbasi
Sajid
S
Malik
Debbie K
DK
Goode
Heather
H
Callaway
Greg
G
Elgar
Esther
E
deGraaff
Javier
J
Lopez-Rios
Rolf
R
Zeller
Karl-Heinz
KH
Grzeschik
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D004742:Enhancer Elements, Genetic; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis / Q000235:genetics*; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics*; D051381:Rats; D023741:Takifugu / Q000235:genetics; D015027:Zebrafish / Q000235:genetics; D000074290:Zinc Finger Protein Gli3
C065683:GLI3 protein, human; D051741:Kruppel-Like Transcription Factors; D009419:Nerve Tissue Proteins; D000074290:Zinc Finger Protein Gli3