IFT20 links kinesin II with a mammalian intraflagellar transport complex that is conserved in motile flagella and sensory cilia

Sheila A Baker; Katie Freeman; Katherine Luby-Phelps; Gregory J Pazour; Joseph C Besharse

doi:10.1074/jbc.M300156200

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IFT20 links kinesin II with a mammalian intraflagellar transport complex that is conserved in motile flagella and sensory cilia

Journal article

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IFT20 links kinesin II with a mammalian intraflagellar transport complex that is conserved in motile flagella and sensory cilia

Sheila A Baker, Katie Freeman, Katherine Luby-Phelps, Gregory J Pazour and Joseph C Besharse

The Journal of biological chemistry, Vol.278(36), pp.34211-34218

09/05/2003

DOI: 10.1074/jbc.M300156200

PMID: 12821668

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Abstract

Intraflagellar transport (IFT) is an evolutionarily conserved mechanism thought to be required for the assembly and maintenance of all eukaryotic cilia and flagella. Although IFT proteins are present in cells with sensory cilia, the organization of IFT protein complexes in those cells has not been analyzed. To determine whether the IFT complex is conserved in the sensory cilia of photo-receptors, we investigated protein interactions among four mammalian IFT proteins: IFT88/Polaris, IFT57/Hippi, IFT52/NGD5, and IFT20. We demonstrate that IFT proteins extracted from bovine photoreceptor outer segments, a modified sensory cilium, co-fractionate at approximately 17 S, similar to IFT proteins extracted from mouse testis. Using antibodies to IFT88 and IFT57, we demonstrate that all four IFT proteins co-immunoprecipitate from lysates of mouse testis, kidney, and retina. We also extended our analysis to interactions outside of the IFT complex and demonstrate an ATP-regulated co-immunoprecipitation of heterotrimeric kinesin II with the IFT complex. The internal architecture of the IFT complex was investigated using the yeast two-hybrid system. IFT20 exhibited a strong interaction with IFT57/Hippi and the kinesin II subunit, KIF3B. Our data indicate that all four mammalian IFT proteins are part of a highly conserved complex in multiple ciliated cell types. Furthermore, IFT20 appears to bridge kinesin II with the IFT complex.

Retina - metabolism

Testis - metabolism

Rod Cell Outer Segment - metabolism

Male

alpha-Galactosidase - metabolism

Kidney - metabolism

Biological Transport

Cattle

Adenosine Triphosphate - metabolism

Flagella - chemistry

Cytoskeletal Proteins - metabolism

Dimerization

Cilia - chemistry

Kinesin

Calcium-Binding Proteins - chemistry

Protein Structure, Tertiary

Ions

Cytoskeletal Proteins - chemistry

Blotting, Western

Precipitin Tests

Hydrolysis

Two-Hybrid System Techniques

Animals

Models, Biological

Photoreceptor Cells - metabolism

Protein Binding

Mice

Muscle Proteins - chemistry

Details

Title: Subtitle: IFT20 links kinesin II with a mammalian intraflagellar transport complex that is conserved in motile flagella and sensory cilia
Creators: Sheila A Baker - Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA
Katie Freeman
Katherine Luby-Phelps
Gregory J Pazour
Joseph C Besharse
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.278(36), pp.34211-34218
DOI: 10.1074/jbc.M300156200
PMID: 12821668
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: EY03222 / NEI NIH HHS R01 EY003222 / NEI NIH HHS GM60992 / NIGMS NIH HHS
Language: English
Date published: 09/05/2003
Academic Unit: Iowa Neuroscience Institute; Biochemistry and Molecular Biology; University College Courses; Ophthalmology and Visual Sciences
Record Identifier: 9984024561902771

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