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PMID:12821668

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Citation

Baker, SA, Freeman, K, Luby-Phelps, K, Pazour, GJ and Besharse, JC (2003) IFT20 links kinesin II with a mammalian intraflagellar transport complex that is conserved in motile flagella and sensory cilia. J. Biol. Chem. 278:34211-8

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.

Links

PubMed Online version:10.1074/jbc.M300156200

Keywords

Adenosine Triphosphate/metabolism; Animals; Biological Transport; Blotting, Western; Calcium-Binding Proteins/chemistry; Cattle; Cilia/chemistry; Cytoskeletal Proteins/chemistry; Cytoskeletal Proteins/metabolism; Dimerization; Flagella/chemistry; Hydrolysis; Ions; Kidney/metabolism; Kinesin; Male; Mice; Models, Biological; Muscle Proteins/chemistry; Photoreceptor Cells/metabolism; Precipitin Tests; Protein Binding; Protein Structure, Tertiary; Retina/metabolism; Rod Cell Outer Segment/metabolism; Testis/metabolism; Two-Hybrid System Techniques; alpha-Galactosidase/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

MOUSE:KIF3A

enables

GO:0120170: intraciliary transport particle B binding

ECO:0000314: direct assay evidence used in manual assertion

F

  • occurs_in:(EMAPA:17373)|occurs_in:(EMAPA:17168)|occurs_in:(EMAPA:17972)

Seeded From UniProt

complete

MOUSE:KIF3A

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:Q61771

F

Seeded From UniProt

complete

MOUSE:KIFA3

enables

GO:0120170: intraciliary transport particle B binding

ECO:0000314: direct assay evidence used in manual assertion

F

  • occurs_in:(EMAPA:17972)|occurs_in:(EMAPA:17168)|occurs_in:(EMAPA:17373)

Seeded From UniProt

complete

MOUSE:IFT20

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(EMAPA:17168)|part_of:(EMAPA:17373)|part_of:(EMAPA:17972)

Seeded From UniProt

complete

MOUSE:IFT20

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:Q8BXG3

F

Seeded From UniProt

complete

MOUSE:IFT20

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:Q61771

F

Seeded From UniProt

complete

MOUSE:IFT88

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(EMAPA:17972)|part_of:(EMAPA:17373)|part_of:(EMAPA:17168)

Seeded From UniProt

complete

MOUSE:KIF3B

enables

GO:0120170: intraciliary transport particle B binding

ECO:0000314: direct assay evidence used in manual assertion

F

  • occurs_in:(EMAPA:17972)|occurs_in:(EMAPA:17373)|occurs_in:(EMAPA:17168)

Seeded From UniProt

complete

MOUSE:KIF3B

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:Q61025

F

Seeded From UniProt

complete

MOUSE:KIF3B

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:P28741

F

Seeded From UniProt

complete

MOUSE:IFT52

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(EMAPA:17373)|part_of:(EMAPA:17168)|part_of:(EMAPA:17972)

Seeded From UniProt

complete

MOUSE:IFT57

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

  • part_of:(EMAPA:17168)|part_of:(EMAPA:17373)|part_of:(EMAPA:17972)

Seeded From UniProt

complete

MOUSE:IFT57

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

PR:Q61025

F

Seeded From UniProt

complete

MOUSE:KIF3A

enables

GO:0120170: intraciliary transport particle B binding

ECO:0000314: direct assay evidence used in manual assertion

F

occurs_in:(EMAPA:17373)|occurs_in:(EMAPA:17168)|occurs_in:(EMAPA:17972)

Seeded From UniProt

complete

MOUSE:IFT88

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(EMAPA:17972)|part_of:(EMAPA:17373)|part_of:(EMAPA:17168)

Seeded From UniProt

complete

MOUSE:IFT57

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

part_of:(EMAPA:17168)|part_of:(EMAPA:17373)|part_of:(EMAPA:17972)

Seeded From UniProt

complete


See also

References

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