Document Detail

The exocyst protein Sec10 is necessary for primary ciliogenesis and cystogenesis in vitro.
MedLine Citation:
PMID:  19297529     Owner:  NLM     Status:  MEDLINE    
Primary cilia are found on many epithelial cell types, including renal tubular epithelial cells, in which they are felt to participate in flow sensing and have been linked to the pathogenesis of cystic renal disorders such as autosomal dominant polycystic kidney disease. We previously localized the exocyst, an eight-protein complex involved in membrane trafficking, to the primary cilium of Madin-Darby canine kidney cells and showed that it was involved in cystogenesis. Here, using short hairpin RNA (shRNA) to knockdown exocyst expression and stable transfection to induce exocyst overexpression, we show that the exocyst protein Sec10 regulates primary ciliogenesis. Using immunofluorescence, scanning, and transmission electron microscopy, primary cilia containing only basal bodies are seen in the Sec10 knockdown cells, and increased ciliogenesis is seen in Sec10-overexpressing cells. These phenotypes do not seem to be because of gross changes in cell polarity, as apical, basolateral, and tight junction proteins remain properly localized. Sec10 knockdown prevents normal cyst morphogenesis when the cells are grown in a collagen matrix, whereas Sec10 overexpression results in increased cystogenesis. Transfection with human Sec10 resistant to the canine shRNA rescues the phenotype, demonstrating specificity. Finally, Par3 was recently shown to regulate primary cilia biogenesis. Par3 and the exocyst colocalized by immunofluorescence and coimmunoprecipitation, consistent with a role for the exocyst in targeting and docking vesicles carrying proteins necessary for primary ciliogenesis.
Xiaofeng Zuo; Wei Guo; Joshua H Lipschutz
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2009-03-18
Journal Detail:
Title:  Molecular biology of the cell     Volume:  20     ISSN:  1939-4586     ISO Abbreviation:  Mol. Biol. Cell     Publication Date:  2009 May 
Date Detail:
Created Date:  2009-05-15     Completed Date:  2009-08-04     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  9201390     Medline TA:  Mol Biol Cell     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2522-9     Citation Subset:  IM    
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
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MeSH Terms
Carrier Proteins / metabolism
Cell Line
Cell Polarity
Cilia / metabolism*,  ultrastructure
Epithelial Cells / cytology,  metabolism,  ultrastructure
Gene Knockdown Techniques
Polycystic Kidney, Autosomal Dominant / metabolism*
Protein Transport
RNA, Small Interfering / metabolism
Tumor Suppressor Proteins / metabolism
Vesicular Transport Proteins / metabolism*
Grant Support
Reg. No./Substance:
0/Carrier Proteins; 0/EXOC5 protein, human; 0/RNA, Small Interfering; 0/Tumor Suppressor Proteins; 0/Vesicular Transport Proteins

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