Abstract
Sonic hedgehog (Shh) is a secreted morphogen that controls the patterning and growth of various tissues in the developing vertebrate embryo, including the central nervous system. Ablation of the FK506-binding protein 38 (FKBP38) gene results in activation of the Shh signaling pathway in mouse embryos, but the molecular mechanism by which FKBP38 suppresses Shh signaling has remained unclear. With the use of a proteomics approach, we have now identified ANKMY2, a protein with three ankyrin repeats and aMYND(myeloid, Nervy, and DEAF- 1)-type Zn2+finger domain, as a molecule that interacts with FKBP38. Co-immunoprecipitation analysis confirmed that endogenous FKBP38 andANKMY2interact in the mouse brain. Depletion or overexpression ofANKMY2resulted in down- and up-regulation of Shh signaling, respectively, in mouse embryonic fibroblasts. Furthermore, combined depletion of both FKBP38 and ANKMY2 attenuated Shh signaling in these cells, suggesting that ANKMY2 acts downstream of FKBP38 to activate the Shh signaling pathway. Targeting of the zebrafish ortholog of mouse Ankmy2 (ankmy2a) in fish embryos with an antisense morpholino oligonucleotide conferred a phenotype reflecting loss of function of the Shh pathway, suggesting that the regulation of Shh signaling by ANKMY2 is conserved between mammals and fish. Our findings thus indicate that the FKBP38-ANKMY2 axis plays a key role in regulation of Shh signaling in vivo.
| Original language | English |
|---|---|
| Pages (from-to) | 25639-25654 |
| Number of pages | 16 |
| Journal | Journal of Biological Chemistry |
| Volume | 289 |
| Issue number | 37 |
| DOIs | |
| Publication status | Published - Sep 12 2014 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Role of the ANKMY2-FKBP38 axis in regulation of the sonic hedgehog (Shh) signaling pathway. / Saita, Shotaro; Shirane, Michiko; Ishitani, Tohru; Shimizu, Nobuyuki; Nakayama, Keiichi.
In: Journal of Biological Chemistry, Vol. 289, No. 37, 12.09.2014, p. 25639-25654.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Role of the ANKMY2-FKBP38 axis in regulation of the sonic hedgehog (Shh) signaling pathway
AU - Saita, Shotaro
AU - Shirane, Michiko
AU - Ishitani, Tohru
AU - Shimizu, Nobuyuki
AU - Nakayama, Keiichi
PY - 2014/9/12
Y1 - 2014/9/12
N2 - Sonic hedgehog (Shh) is a secreted morphogen that controls the patterning and growth of various tissues in the developing vertebrate embryo, including the central nervous system. Ablation of the FK506-binding protein 38 (FKBP38) gene results in activation of the Shh signaling pathway in mouse embryos, but the molecular mechanism by which FKBP38 suppresses Shh signaling has remained unclear. With the use of a proteomics approach, we have now identified ANKMY2, a protein with three ankyrin repeats and aMYND(myeloid, Nervy, and DEAF- 1)-type Zn2+finger domain, as a molecule that interacts with FKBP38. Co-immunoprecipitation analysis confirmed that endogenous FKBP38 andANKMY2interact in the mouse brain. Depletion or overexpression ofANKMY2resulted in down- and up-regulation of Shh signaling, respectively, in mouse embryonic fibroblasts. Furthermore, combined depletion of both FKBP38 and ANKMY2 attenuated Shh signaling in these cells, suggesting that ANKMY2 acts downstream of FKBP38 to activate the Shh signaling pathway. Targeting of the zebrafish ortholog of mouse Ankmy2 (ankmy2a) in fish embryos with an antisense morpholino oligonucleotide conferred a phenotype reflecting loss of function of the Shh pathway, suggesting that the regulation of Shh signaling by ANKMY2 is conserved between mammals and fish. Our findings thus indicate that the FKBP38-ANKMY2 axis plays a key role in regulation of Shh signaling in vivo.
AB - Sonic hedgehog (Shh) is a secreted morphogen that controls the patterning and growth of various tissues in the developing vertebrate embryo, including the central nervous system. Ablation of the FK506-binding protein 38 (FKBP38) gene results in activation of the Shh signaling pathway in mouse embryos, but the molecular mechanism by which FKBP38 suppresses Shh signaling has remained unclear. With the use of a proteomics approach, we have now identified ANKMY2, a protein with three ankyrin repeats and aMYND(myeloid, Nervy, and DEAF- 1)-type Zn2+finger domain, as a molecule that interacts with FKBP38. Co-immunoprecipitation analysis confirmed that endogenous FKBP38 andANKMY2interact in the mouse brain. Depletion or overexpression ofANKMY2resulted in down- and up-regulation of Shh signaling, respectively, in mouse embryonic fibroblasts. Furthermore, combined depletion of both FKBP38 and ANKMY2 attenuated Shh signaling in these cells, suggesting that ANKMY2 acts downstream of FKBP38 to activate the Shh signaling pathway. Targeting of the zebrafish ortholog of mouse Ankmy2 (ankmy2a) in fish embryos with an antisense morpholino oligonucleotide conferred a phenotype reflecting loss of function of the Shh pathway, suggesting that the regulation of Shh signaling by ANKMY2 is conserved between mammals and fish. Our findings thus indicate that the FKBP38-ANKMY2 axis plays a key role in regulation of Shh signaling in vivo.
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UR - http://www.scopus.com/inward/citedby.url?scp=84907164686&partnerID=8YFLogxK
U2 - 10.1074/jbc.M114.558635
DO - 10.1074/jbc.M114.558635
M3 - Article
C2 - 25077969
AN - SCOPUS:84907164686
VL - 289
SP - 25639
EP - 25654
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 37
ER -