Direct Interactions of Intraflagellar Transport Complex B Proteins IFT88, IFT52, and IFT46

Ben F Lucker; Mark S Miller; Slawomir A Dziedzic; Philip T Blackmarr; Douglas G Cole

doi:10.1074/jbc.M110.106997

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Direct Interactions of Intraflagellar Transport Complex B Proteins IFT88, IFT52, and IFT46

Journal article

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Direct Interactions of Intraflagellar Transport Complex B Proteins IFT88, IFT52, and IFT46

Ben F Lucker, Mark S Miller, Slawomir A Dziedzic, Philip T Blackmarr and Douglas G Cole

The Journal of biological chemistry, Vol.285(28), pp.21508-21518

07/09/2010

DOI: https://doi.org/10.1074/jbc.M110.106997

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https://hdl.handle.net/2376/113327

PMCID: PMC2898376

PMID: 20435895

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Abstract

Molecular Motors

Flagella

Intraflagellar Transport

Kinesin

Chlamydomonas

Cell Biology

Protein Assembly

Protein Cross-linking

Protein-Protein Interactions

Cilia

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 ∼500-kDa core that contains IFT88, 2× IFT81, 2× 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.

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Title: Direct Interactions of Intraflagellar Transport Complex B Proteins IFT88, IFT52, and IFT46
Creators: Ben F Lucker - From the Department of Microbiology, Molecular Biology, and Biochemistry and the Center for Reproductive Biology, University of Idaho, Moscow, Idaho 83844
Mark S Miller - From the Department of Microbiology, Molecular Biology, and Biochemistry and the Center for Reproductive Biology, University of Idaho, Moscow, Idaho 83844
Slawomir A Dziedzic - From the Department of Microbiology, Molecular Biology, and Biochemistry and the Center for Reproductive Biology, University of Idaho, Moscow, Idaho 83844
Philip T Blackmarr - From the Department of Microbiology, Molecular Biology, and Biochemistry and the Center for Reproductive Biology, University of Idaho, Moscow, Idaho 83844
Douglas G Cole - From the Department of Microbiology, Molecular Biology, and Biochemistry and the Center for Reproductive Biology, University of Idaho, Moscow, Idaho 83844
Publication Details: The Journal of biological chemistry, Vol.285(28), pp.21508-21518
Academic Unit: Center for Reproductive Biology
Publisher: American Society for Biochemistry and Molecular Biology; 9650 Rockville Pike, Bethesda, MD 20814, U.S.A
Grant note: R01-GM61920; P20-RR016454 / National Institutes of Health
Identifiers: 99900547544301842
Language: English
Resource Type: Journal article