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Krüppel-like zinc finger transcription factors compose the largest transcription factor family in the mammalian genome. However, the functions for the majority of these transcription factors as well as their in vivo downstream targets are not clear. We have functionally characterized a novel KRAB domain zinc finger transcription factor ZNF431 using both in vitro and in vivo assays. ZNF431 is a nuclear transcriptional repressor whose repressive activity depends on its association with HDAC1 and -2. Using the limbmesenchymal cell line MPLB, we identified Patched1 as a direct transcriptional target of ZNF431. Promoter analyses revealed three ZNF431 binding sites that bind to ZNF431 both in vitro and in vivo as revealed by gel-shift and chromatin immunoprecipitation, respectively. Mutations of these three sites abolished ZNF431 repression in transient transfection assays. Moreover, overexpressing ZNF431 in MPLB cells or in Xenopus and mouse embryos strongly repressed Patched1 expression. On the other hand, shRNA knockdown of ZNF431 in MPLB cells elevated Patched1 expression. Finally, hedgehog signaling readout was reduced in ZNF431 overexpression but elevated in ZNF431 knockdown MPLB cells. Our results indicate that ZNF431 directly represses Patched1 expression and likely functions to repress the hedgehog response in cells.
FIGURE 6. Overexpression of ZNF431 repressed Ptch1 expression in Xenopus
and transgenic mouse embryos. A, shown are whole mount in situ
hybridization results of Xptc1 expression. The injected halves (Inj.) are indicated
by X-gal staining. B, a schematic diagram of the ZNF431 transgene is
shown. HA-tagged ZNF431 was driven by the chicken -actin promoter
preceded by a LoxP-flanked transcriptional stop cassette and a -globin
intron. The relative positions of genotyping primers are indicated by arrows.
C, whole mount in situ hybridization of Ptch1 expression in E10.5 wild-type
(upper) or HA-ZNF431 transgenic embryos (lower) is show. Ptch1 staining in
the limb bud is indicated by arrows. D, real-time PCR quantification of Ptch1
expression in wild-type or HA-ZNF431 transgenic embryos is shown. The
results were shown as relative Ptch1 expression. Tg, transgenic.
Agren,
Expression of the PTCH1 tumor suppressor gene is regulated by alternative promoters and a single functional Gli-binding site.
2004, Pubmed
Agren,
Expression of the PTCH1 tumor suppressor gene is regulated by alternative promoters and a single functional Gli-binding site.
2004,
Pubmed Ahn,
In vivo analysis of quiescent adult neural stem cells responding to Sonic hedgehog.
2005,
Pubmed Ahringer,
NuRD and SIN3 histone deacetylase complexes in development.
2000,
Pubmed Bellefroid,
The human genome contains hundreds of genes coding for finger proteins of the Krüppel type.
1989,
Pubmed Bellefroid,
The evolutionarily conserved Krüppel-associated box domain defines a subfamily of eukaryotic multifingered proteins.
1991,
Pubmed
,
Xenbase Ding,
SysZNF: the C2H2 zinc finger gene database.
2009,
Pubmed Fasano,
shRNA knockdown of Bmi-1 reveals a critical role for p21-Rb pathway in NSC self-renewal during development.
2007,
Pubmed Fried,
Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis.
1981,
Pubmed Friedman,
KAP-1, a novel corepressor for the highly conserved KRAB repression domain.
1996,
Pubmed Gebelein,
Sequence-specific transcriptional repression by KS1, a multiple-zinc-finger-Krüppel-associated box protein.
2001,
Pubmed Goodrich,
Conservation of the hedgehog/patched signaling pathway from flies to mice: induction of a mouse patched gene by Hedgehog.
1996,
Pubmed Groner,
KRAB-zinc finger proteins and KAP1 can mediate long-range transcriptional repression through heterochromatin spreading.
2010,
Pubmed Hardcastle,
The Shh signalling pathway in tooth development: defects in Gli2 and Gli3 mutants.
1998,
Pubmed
,
Xenbase Harland,
In situ hybridization: an improved whole-mount method for Xenopus embryos.
1991,
Pubmed
,
Xenbase Helms,
Sonic hedgehog participates in craniofacial morphogenesis and is down-regulated by teratogenic doses of retinoic acid.
1997,
Pubmed Hennemann,
Identification of a novel Krüppel-associated box domain protein, Krim-1, that interacts with c-Myc and inhibits its oncogenic activity.
2003,
Pubmed Hering,
Characterization and chondrocyte differentiation stage-specific expression of KRAB zinc-finger protein gene ZNF470.
2004,
Pubmed Ingham,
Hedgehog signaling in animal development: paradigms and principles.
2001,
Pubmed Jheon,
Characterization of a novel KRAB/C2H2 zinc finger transcription factor involved in bone development.
2001,
Pubmed Kim,
A novel member of the RING finger family, KRIP-1, associates with the KRAB-A transcriptional repressor domain of zinc finger proteins.
1996,
Pubmed Kovacs,
Arrestin development: emerging roles for beta-arrestins in developmental signaling pathways.
2009,
Pubmed Lin,
Temporal and spatial dissection of Shh signaling in genital tubercle development.
2009,
Pubmed Lum,
The Hedgehog response network: sensors, switches, and routers.
2004,
Pubmed Matsuda,
Targeting of Krüppel-associated box-containing zinc finger proteins to centromeric heterochromatin. Implication for the gene silencing mechanisms.
2001,
Pubmed Miller,
An improved zinc-finger nuclease architecture for highly specific genome editing.
2007,
Pubmed Moosmann,
Transcriptional repression by RING finger protein TIF1 beta that interacts with the KRAB repressor domain of KOX1.
1996,
Pubmed Nagao,
Identification and characterization of multiple isoforms of a murine and human tumor suppressor, patched, having distinct first exons.
2005,
Pubmed Niswander,
A positive feedback loop coordinates growth and patterning in the vertebrate limb.
1994,
Pubmed Pabo,
Design and selection of novel Cys2His2 zinc finger proteins.
2001,
Pubmed Pan,
gamma-secretase functions through Notch signaling to maintain skin appendages but is not required for their patterning or initial morphogenesis.
2004,
Pubmed Pavletich,
Zinc finger-DNA recognition: crystal structure of a Zif268-DNA complex at 2.1 A.
1991,
Pubmed Schultz,
Targeting histone deacetylase complexes via KRAB-zinc finger proteins: the PHD and bromodomains of KAP-1 form a cooperative unit that recruits a novel isoform of the Mi-2alpha subunit of NuRD.
2001,
Pubmed Seo,
Geminin regulates neuronal differentiation by antagonizing Brg1 activity.
2005,
Pubmed
,
Xenbase Sinha,
Purmorphamine activates the Hedgehog pathway by targeting Smoothened.
2006,
Pubmed Solomon,
Mapping protein-DNA interactions in vivo with formaldehyde: evidence that histone H4 is retained on a highly transcribed gene.
1988,
Pubmed St-Jacques,
Sonic hedgehog signaling is essential for hair development.
1998,
Pubmed Treviño,
MPLB-2: a posterior signaling cell line derived from the mouse limb bud.
1993,
Pubmed Urrutia,
KRAB-containing zinc-finger repressor proteins.
2003,
Pubmed Vissing,
Repression of transcriptional activity by heterologous KRAB domains present in zinc finger proteins.
1995,
Pubmed Yoon,
Gene expression profiling leads to identification of GLI1-binding elements in target genes and a role for multiple downstream pathways in GLI1-induced cell transformation.
2002,
Pubmed
