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PMID: 19253336 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Characterization of mouse IFT complex B.

Cell motility and the cytoskeleton ·Vol. 66 ·No. 8 ·2009-08-00 ·Pages 457-68

Follit JA, Xu F, Keady BT, Pazour GJ

Abstract

The primary cilium plays a key role in the development of mammals and in the maintenance of health. Primary cilia are assembled and maintained by the process of intraflagellar transport (IFT). In this work, we characterize mouse IFT complex B by identifying all of the mammalian orthologues of complex B and B-associated proteins previously identified in Chlamydomonas and Caenorhabditis and also identify a new component (IFT25/Hspb11) of complex B by database analysis. We tagged each of these proteins with the FLAG epitope and show that all except IFT172 and IFT20 localize to cilia and the peri-basal body or centrosomal region at the base of cilia. All of the proteins except IFT172 immunoprecipitate IFT88 indicating that they are co-assembled into a complex. IFT20 is the only complex B protein that localizes to the Golgi apparatus. However, overexpression of IFT54/Traf3ip1, the mouse orthologue of Dyf-11/Elipsa, displaces IFT20 from the Golgi apparatus. IFT54 does not localize to the Golgi complex nor does it interact with GMAP210, which is the protein that anchors IFT20 to the Golgi apparatus. This suggests that IFT54s effect on IFT20 is a dominant negative phenotype caused by its overexpression. Cell Motil. Cytoskeleton 2009. (c) 2009 Wiley-Liss, Inc.

MeSH Terms
Adaptor Proteins, Signal Transducing Animals Blotting, Western Carrier Proteins/metabolism Cell Line Cilia/metabolism Cytoskeletal Proteins Flagella/metabolism Golgi Apparatus/metabolism Immunoprecipitation Intracellular Signaling Peptides and Proteins/metabolism Mice Nuclear Proteins/metabolism Protein Binding Tumor Suppressor Proteins/metabolism
Chemicals
Adaptor Proteins, Signal Transducing Carrier Proteins Cytoskeletal Proteins Ift172 protein, mouse Ift20 protein, mouse Intracellular Signaling Peptides and Proteins Nuclear Proteins TRIP11 protein, mouse Tg737Rpw protein, mouse Tumor Suppressor Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Follit John A
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Xu Fenghui
Keady Brian T
Pazour Gregory J
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Article Info
Journal
Cell motility and the cytoskeleton
Abbr.
Cell Motil Cytoskeleton
ISSN
1097-0169
DOI
10.1002/cm.20346
Published
2009-08-00
Pages
457-68
Language
English
Region
United States
NLM ID
8605339
PMCID
PMC2753169
Subset
IM
Grants
NIDDK NIH HHS · P30 DK032520 · United States
NIGMS NIH HHS · R01 GM060992-09 · United States
NIGMS NIH HHS · R01 GM060992 · United States
NIGMS NIH HHS · GM060992 · United States
NIGMS NIH HHS · R01 GM060992-08 · United States
External Links
PubMed source DOI: 10.1002/cm.20346 PMC Full Text
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