Document Detail


Subunit interactions and organization of the Chlamydomonas reinhardtii intraflagellar transport complex A proteins.
MedLine Citation:
PMID:  22170070     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chlamydomonas reinhardtii intraflagellar transport (IFT) particles can be biochemically resolved into two smaller assemblies, complexes A and B, that contain up to six and 15 protein subunits, respectively. We provide here the proteomic and immunological analyses that verify the identity of all six Chlamydomonas A proteins. Using sucrose density gradient centrifugation and antibody pulldowns, we show that all six A subunits are associated in a 16 S complex in both the cell bodies and flagella. A significant fraction of the cell body IFT43, however, exhibits a much slower sedimentation of ∼2 S and is not associated with the IFT A complex. To identify interactions between the six A proteins, we combined exhaustive yeast-based two-hybrid analysis, heterologous recombinant protein expression in Escherichia coli, and analysis of the newly identified complex A mutants, ift121 and ift122. We show that IFT121 and IFT43 interact directly and provide evidence for additional interactions between IFT121 and IFT139, IFT121 and IFT122, IFT140 and IFT122, and IFT140 and IFT144. The mutant analysis further allows us to propose that a subset of complex A proteins, IFT144/140/122, can form a stable 12 S subcomplex that we refer to as the IFT A core. Based on these results, we propose a model for the spatial arrangement of the six IFT A components.
Authors:
Robert H Behal; Mark S Miller; Hongmin Qin; Ben F Lucker; Alexis Jones; Douglas G Cole
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2011-12-14
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  287     ISSN:  1083-351X     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2012 Apr 
Date Detail:
Created Date:  2012-04-11     Completed Date:  2012-06-06     Revised Date:  2013-06-27    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  11689-703     Citation Subset:  IM    
Affiliation:
Department of Biological Sciences and Center for Reproductive Biology, University of Idaho, Moscow, Idaho 83844, USA.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AY686103;  EF586682;  EF592032;  EF592033;  EF599100;  JQ361074
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Biological Transport
Chlamydomonas reinhardtii / genetics,  metabolism*
Escherichia coli
Flagella / chemistry,  metabolism*
Gene Knockout Techniques
Immunoprecipitation
Models, Molecular
Molecular Sequence Data
Multiprotein Complexes / chemistry,  genetics,  metabolism
Phylogeny
Plant Proteins / chemistry,  genetics,  metabolism*
Protein Binding
Protein Interaction Domains and Motifs
Protein Stability
Protein Structure, Quaternary
Protein Subunits / chemistry,  metabolism
Recombinant Proteins / chemistry,  metabolism
Two-Hybrid System Techniques
Grant Support
ID/Acronym/Agency:
P20-RR016454/RR/NCRR NIH HHS; R01-GM61920/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Multiprotein Complexes; 0/Plant Proteins; 0/Protein Subunits; 0/Recombinant Proteins
Comments/Corrections

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