Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Subscribe

Sci. Signal., 30 September 2008
Vol. 1, Issue 39, p. ra7
[DOI: 10.1126/scisignal.1162925]

RESEARCH ARTICLES

Kinome siRNA Screen Identifies Regulators of Ciliogenesis and Hedgehog Signal Transduction

Marie Evangelista1*, Tze Yang Lim1, James Lee1, Leon Parker2, Amir Ashique1, Andrew S. Peterson1, Weilan Ye2, David P. Davis1, and Frederic J. de Sauvage1*

1 Department of Molecular Biology, Genentech, South San Francisco, CA 94080, USA.
2 Department of Tumor Biology and Angiogenesis (TBA), Genentech, South San Francisco, CA 94080, USA.

Abstract: Disruption or improper activation of the Hedgehog (Hh) pathway is associated with developmental abnormalities and cancer. Although characterized in Drosophila, the mechanisms that mediate the Hh signal downstream of the Smoothened (Smo) seven-transmembrane protein in vertebrates remain poorly understood. In particular, the Fused (Fu) kinase, which mediates Hh signaling in flies, is dispensable in mammals. To identify kinases that positively regulate the Hh pathway in mammals, we screened a mouse kinome small interfering RNA library and validated nine candidates that modulate Hh signaling. Among these candidates, Nek1 and Prkra did not directly function in the Hh pathway but exerted their effects on Hh signaling indirectly through a primary role in ciliogenesis. In contrast, another kinase, Cdc2l1, directly participated in the Hh pathway. Cdc2l1 was necessary and sufficient for activation of the Hh pathway, functioning downstream of Smo and upstream of the Glioma-associated (Gli) transcription factors. More specifically, Cdc2l1 interacted with the negative regulator Suppressor of Fused (Sufu) and relieved its inhibition on Gli, thus providing a mechanism for how Cdc2l1 might play a role in Hh signaling. Finally, with zebrafish as model organism, we showed that Cdc2l1 activated the Hh pathway in vivo. We propose that Cdc2l1 is a previously unrecognized member of the Hh signal transduction cascade.

* To whom correspondence should be addressed. E-mail: evangelm{at}gene.com (M.E.) and sauvage{at}gene.com (F.J.d.S.)

Citation: M. Evangelista, T. Y. Lim, J. Lee, L. Parker, A. Ashique, A. S. Peterson, W. Ye, D. P. Davis, F. J. de Sauvage, Kinome siRNA Screen Identifies Regulators of Ciliogenesis and Hedgehog Signal Transduction. Sci. Signal. 1, ra7 (2008).

Read the Full Text

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Rer1p maintains ciliary length and signaling by regulating {gamma}-secretase activity and Foxj1a levels.
N. Jurisch-Yaksi, A. J. Rose, H. Lu, T. Raemaekers, S. Munck, P. Baatsen, V. Baert, W. Vermeire, S. J. Scales, D. Verleyen, et al. (2013)
J. Cell Biol. 200, 709-720
   Abstract »    Full Text »    PDF »
Systematic Kinome shRNA Screening Identifies CDK11 (PITSLRE) Kinase Expression Is Critical for Osteosarcoma Cell Growth and Proliferation.
Z. Duan, J. Zhang, E. Choy, D. Harmon, X. Liu, P. Nielsen, H. Mankin, N. S. Gray, and F. J. Hornicek (2012)
Clin. Cancer Res. 18, 4580-4588
   Abstract »    Full Text »    PDF »
Proteomic and Functional Genomic Landscape of Receptor Tyrosine Kinase and Ras to Extracellular Signal-Regulated Kinase Signaling.
A. A. Friedman, G. Tucker, R. Singh, D. Yan, A. Vinayagam, Y. Hu, R. Binari, P. Hong, X. Sun, M. Porto, et al. (2011)
Science Signaling 4, rs10
   Abstract »    Full Text »    PDF »
Functional characterization of putative cilia genes by high-content analysis.
C. K. Lai, N. Gupta, X. Wen, L. Rangell, B. Chih, A. S. Peterson, J. F. Bazan, L. Li, and S. J. Scales (2011)
Mol. Biol. Cell 22, 1104-1119
   Abstract »    Full Text »    PDF »
Genome-Wide RNAi Screen Reveals Disease-Associated Genes That Are Common to Hedgehog and Wnt Signaling.
L. S. Jacob, X. Wu, M. E. Dodge, C.-W. Fan, O. Kulak, B. Chen, W. Tang, B. Wang, J. F. Amatruda, and L. Lum (2011)
Science Signaling 4, ra4
   Abstract »    Full Text »    PDF »
The RNA Binding Motif Protein 15B (RBM15B/OTT3) Is a Functional Competitor of Serine-Arginine (SR) Proteins and Antagonizes the Positive Effect of the CDK11p110-Cyclin L2{alpha} Complex on Splicing.
P. Loyer, A. Busson, J. H. Trembley, J. Hyle, J. Grenet, W. Zhao, C. Ribault, T. Montier, V. J. Kidd, and J. M. Lahti (2011)
J. Biol. Chem. 286, 147-159
   Abstract »    Full Text »    PDF »
Dual Function of UNC-51-like Kinase 3 (Ulk3) in the Sonic Hedgehog Signaling Pathway.
A. Maloverjan, M. Piirsoo, L. Kasak, L. Peil, T. Osterlund, and P. Kogerman (2010)
J. Biol. Chem. 285, 30079-30090
   Abstract »    Full Text »    PDF »
Mechanism and evolution of cytosolic Hedgehog signal transduction.
C. W. Wilson and P.-T. Chuang (2010)
Development 137, 2079-2094
   Abstract »    Full Text »    PDF »
Kinetics of Hedgehog-Dependent Full-Length Gli3 Accumulation in Primary Cilia and Subsequent Degradation.
X. Wen, C. K. Lai, M. Evangelista, J.-A. Hongo, F. J. de Sauvage, and S. J. Scales (2010)
Mol. Cell. Biol. 30, 1910-1922
   Abstract »    Full Text »    PDF »
Variations in Hedgehog signaling: divergence and perpetuation in Sufu regulation of Gli.
L. Ruel and P. P. Therond (2009)
Genes & Dev. 23, 1843-1848
   Abstract »    Full Text »    PDF »
Mouse hitchhiker mutants have spina bifida, dorso-ventral patterning defects and polydactyly: identification of Tulp3 as a novel negative regulator of the Sonic hedgehog pathway.
V. L. Patterson, C. Damrau, A. Paudyal, B. Reeve, D. T. Grimes, M. E. Stewart, D. J. Williams, P. Siggers, A. Greenfield, and J. N. Murdoch (2009)
Hum. Mol. Genet. 18, 1719-1739
   Abstract »    Full Text »    PDF »
Science Signaling Podcast: 07 April 2009.
B. D. Manning and A. M. VanHook (2009)
Science Signaling 2, pc7
   Abstract »    Full Text »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882