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

Research output: Contribution to journalArticle

98 Citations (Scopus)

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 photoreceptors, 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 ∼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.

Original languageEnglish (US)
Pages (from-to)34211-34218
Number of pages8
JournalJournal of Biological Chemistry
Volume278
Issue number36
DOIs
StatePublished - Sep 5 2003

Fingerprint

Flagella
Cilia
Carrier Proteins
Testis
Two-Hybrid System Techniques
kinesin-II
Hybrid systems
Immunoprecipitation
Yeast
Retina
Adenosine Triphosphate
Maintenance
Kidney
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

IFT20 Links Kinesin II with a Mammalian Intraflagellar Transport Complex that Is Conserved in Motile Flagella and Sensory Cilia. / Baker, Sheila A.; Freeman, Katie; Luby-Phelps, Katherine; Pazour, Gregory J.; Besharse, Joseph C.

In: Journal of Biological Chemistry, Vol. 278, No. 36, 05.09.2003, p. 34211-34218.

Research output: Contribution to journalArticle

Baker, Sheila A. ; Freeman, Katie ; Luby-Phelps, Katherine ; Pazour, Gregory J. ; Besharse, Joseph C. / IFT20 Links Kinesin II with a Mammalian Intraflagellar Transport Complex that Is Conserved in Motile Flagella and Sensory Cilia. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 36. pp. 34211-34218.
@article{8eab375a748e4b1e81773105a91cf984,
title = "IFT20 Links Kinesin II with a Mammalian Intraflagellar Transport Complex that Is Conserved in Motile Flagella and Sensory Cilia",
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 photoreceptors, 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 ∼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.",
author = "Baker, {Sheila A.} and Katie Freeman and Katherine Luby-Phelps and Pazour, {Gregory J.} and Besharse, {Joseph C.}",
year = "2003",
month = "9",
day = "5",
doi = "10.1074/jbc.M300156200",
language = "English (US)",
volume = "278",
pages = "34211--34218",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "36",

}

TY - JOUR

T1 - IFT20 Links Kinesin II with a Mammalian Intraflagellar Transport Complex that Is Conserved in Motile Flagella and Sensory Cilia

AU - Baker, Sheila A.

AU - Freeman, Katie

AU - Luby-Phelps, Katherine

AU - Pazour, Gregory J.

AU - Besharse, Joseph C.

PY - 2003/9/5

Y1 - 2003/9/5

N2 - 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 photoreceptors, 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 ∼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.

AB - 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 photoreceptors, 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 ∼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.

UR - http://www.scopus.com/inward/record.url?scp=0141706698&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0141706698&partnerID=8YFLogxK

U2 - 10.1074/jbc.M300156200

DO - 10.1074/jbc.M300156200

M3 - Article

VL - 278

SP - 34211

EP - 34218

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 36

ER -