Evc2 is a positive modulator of Hedgehog signalling that interacts with Evc at the cilia membrane and is also found in the nucleus

BMC Biology

Research Article

Katie MacArthur¹  Helen J Blair¹  Yu-Ning Liu¹  Jennifer Campbell¹  Judith A Goodship¹  Stuart Tompson²  Victor L Ruiz-Perez³  Chris P Ponting⁴ 

[1] Institute of Human Genetics, Newcastle University, Centre for Life, Central Parkway, NE1 3BZ, Newcastle Upon Tyne, UK;Institute of Human Genetics, Newcastle University, Centre for Life, Central Parkway, NE1 3BZ, Newcastle Upon Tyne, UK;Medical Genetics Institute, Cedars-Sinai Medical Center 8727 West Third Street, Suite 203, 90048, Los Angeles, CA, USA;Instituto de Investigaciones Biomédicas de Madrid (CSIC-UAM) and Ciber de enfermedades raras (Ciberer), Arturo Duperier, 4, 28029, Madrid, Spain;MRC Functional Genomics Unit, University of Oxford, Department of Physiology, Anatomy and Genetics, South Parks Road, OX1 3QX, Oxford, UK;

关键词: Basal Body;    Primary Cilium;    Cartilage Growth Plate;    MC3T3 Cell;    IMCD3 Cell;   

DOI  :  10.1186/1741-7007-9-14

 received in 2011-02-04, accepted in 2011-02-28,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundEvc is essential for Indian Hedgehog (Hh) signalling in the cartilage growth plate. The gene encoding Evc2 is in close proximity in divergent orientation to Evc and mutations in both human genes lead to the chondrodysplasia Ellis-van Creveld syndrome.ResultsBioinformatic analysis reveals that the Evc and Evc2 genes arose through a duplication event early in metazoan evolution and were subsequently lost in arthropods and nematodes. Here we demonstrate that Evc2 is essential for Hh pathway activation in response to the Smo agonist purmorphamine. A yeast two-hybrid screen using Evc as bait identified Evc2 as an Evc binding partner and we confirmed the interaction by immunoprecipitation. We developed anti-Evc2 antibodies and show that Evc2 and Evc co-localize at the basal body and also on primary cilia. In transfected cells, basal body and cilia localization is observed when Evc and Evc2 constructs are co-transfected but not when either construct is transfected individually. We show that Evc and Evc2 are cilia transmembrane proteins, the C-terminus for both being intracellular and Evc2, but not Evc, having an extracellular portion. Furthermore, Evc is absent at the basal body in Evc2 null cells. Using Western blots of cytoplasmic and nuclear protein, we also demonstrate that full length Evc2 but not Evc, is located in the nucleus.ConclusionsWe demonstrate for the first time that Evc2 is a positive regulator of the Hh signalling pathway and that it is located at the basal body of primary cilia. We show that the presence of Evc and Evc2 at the basal body and cilia membrane is co-dependent. In addition, Evc2, but not Evc, is present in the cell nucleus suggesting movement of Evc2 between the cilium and nucleus.

【 授权许可】

Unknown   
© Blair et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.