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PMID:20435895

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Citation

Lucker, BF, Miller, MS, Dziedzic, SA, Blackmarr, PT and Cole, DG (2010) Direct interactions of intraflagellar transport complex B proteins IFT88, IFT52, and IFT46. J. Biol. Chem. 285:21508-18

Abstract

Intraflagellar transport (IFT) particles of Chlamydomonas reinhardtii contain two distinct protein complexes, A and B, composed of at least 6 and 15 protein subunits, respectively. As isolated from C. reinhardtii flagella, IFT complex B can be further reduced to a approximately 500-kDa core that contains IFT88, 2x IFT81, 2x IFT74/72, IFT52, IFT46, IFT27, IFT25, and IFT22. In this study, yeast-based two-hybrid analysis was combined with bacterial coexpression to show that three of the core B subunits, IFT88, IFT52, and IFT46, interact directly with each other and, together, are capable of forming a ternary complex. Chemical cross-linking results support the IFT52-IFT88 interaction and provide additional evidence of an association between IFT27 and IFT81. With previous studies showing that IFT81 and IFT74/72 interact to form a (IFT81)(2)(IFT74/72)(2) heterotetramer and that IFT27 and IFT25 form a heterodimer, the architecture of complex B is revealing itself. Last, electroporation of recombinant IFT46 was used to rescue flagellar assembly of a newly identified ift46 mutant and to monitor in vivo localization and movement of the IFT particles.

Links

PubMed PMC2898376 Online version:10.1074/jbc.M110.106997

Keywords

Algal Proteins/metabolism; Biological Transport; Carrier Proteins/metabolism; Chlamydomonas reinhardtii/genetics; Chlamydomonas reinhardtii/metabolism; Cross-Linking Reagents/chemistry; Electroporation; Flagella/metabolism; Gene Expression Regulation; Models, Biological; Mutagenesis; Phenotype; Plasmids/metabolism; Protein Binding; Protozoan Proteins/metabolism; Recombinant Proteins/chemistry; Two-Hybrid System Techniques

Significance

Annotations

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

CHLRE:IFT46

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9FPW0

F

Seeded From UniProt

complete

CHLRE:IFT46

located_in

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT27

located_in

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT27

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT81

located_in

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT81

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT74

located_in

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT74

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:Q946G4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9FPW0

F

Seeded From UniProt

complete

CHLRE:Q946G4

enables

GO:0008022: protein C-terminus binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:A2T2X4

F

Seeded From UniProt

complete

CHLRE:Q946G4

located_in

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:Q946G4

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:Q9FPW0

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q946G4

F

Seeded From UniProt

complete

CHLRE:Q9FPW0

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:A2T2X4

F

Seeded From UniProt

complete

CHLRE:Q9FPW0

located_in

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:Q9FPW0

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:Q6RCE1

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:Q6RCE1

part_of

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT46

part_of

GO:0030992: intraciliary transport particle B

ECO:0000304: author statement supported by traceable reference used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT46

enables

GO:0008022: protein C-terminus binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q946G4

F

Seeded From UniProt

complete

CHLRE:IFT46

part_of

GO:0030992: intraciliary transport particle B

ECO:0000305: curator inference used in manual assertion

GO:0031514

C

Seeded From UniProt

complete

CHLRE:IFT46

part_of

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT46

colocalizes_with

GO:0036064: ciliary basal body

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:IFT46

involved_in

GO:0042073: intraciliary transport

ECO:0000305: curator inference used in manual assertion

GO:0060271

P

Seeded From UniProt

complete

CHLRE:IFT46

involved_in

GO:0060271: cilium assembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

CHLRE:IFT46

involved_in

GO:0050821: protein stabilization

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

CHLRE:Q68RJ5

part_of

GO:0030992: intraciliary transport particle B

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CHLRE:Q68RJ5

part_of

GO:0031514: motile cilium

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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