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PMID: 15563613 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S.

Dimeric novel HSP40 is incorporated into the radial spoke complex during the assembly process in flagella.

Molecular biology of the cell ·Vol. 16 ·No. 2 ·2005-02-00 ·Pages 637-48

Yang C, Compton MM, Yang P

Abstract

The radial spoke is a stable structural complex in the 9 + 2 axoneme for the control of flagellar motility. However, the spokes in Chlamydomonas mutant pf24 are heterogeneous and unstable, whereas several spoke proteins are reduced differentially. To elucidate the defective mechanism, we clone RSP16, a prominent spoke protein diminished in pf24 axonemes. Unexpectedly, RSP16 is a novel HSP40 member of the DnaJ superfamily that assists chaperones in various protein-folding-related processes. Importantly, RSP16 is uniquely excluded from the 12S spoke precursor complex that is packaged in the cell body and transported toward the flagellar tip to be converted into mature 20S axonemal spokes. Rather, RSP16, transported separately, joins the precursor complex in flagella. Furthermore, RSP16 molecules in vitro and in flagella form homodimers, a characteristic required for the cochaperone activity of HSP40. We postulate that the spoke HSP40 operates as a cochaperone to assist chaperone machinery at the flagellar tip to actively convert the smaller spoke precursor and itself into the mature stable complex; failure of the interaction between the spoke HSP40 and its target polypeptide results in heterogeneous unstable radial spokes in pf24.

MeSH Terms
Amino Acid Sequence Animals Blotting, Western Cell Fractionation Chlamydomonas/chemistry,genetics,metabolism Chromatography, Affinity Chromatography, Liquid Chromosome Mapping Cloning, Molecular Dimerization Electrophoresis, Gel, Two-Dimensional Electrophoresis, Polyacrylamide Gel Flagella/chemistry,metabolism HSP40 Heat-Shock Proteins Heat-Shock Proteins/chemistry,metabolism Molecular Sequence Data Mutation Phylogeny Protein Structure, Tertiary Protozoan Proteins/genetics,metabolism Recombinant Proteins/metabolism Reverse Transcriptase Polymerase Chain Reaction Sequence Homology, Amino Acid
Chemicals
HSP40 Heat-Shock Proteins Heat-Shock Proteins Protozoan Proteins Recombinant Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Yang Chun
Department of Biological Sciences, Marquette University, Milwaukee, WI 53233, USA.
Compton Mark M
Yang Pinfen
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e04-09-0787
Published
2005-02-00
Epub
2004-00-24
Pages
637-48
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC545900
Subset
IM
Grants
NIGMS NIH HHS · GM68101 · United States
NIGMS NIH HHS · R01 GM068101-02 · United States
NIGMS NIH HHS · R01 GM068101-03 · United States
NIGMS NIH HHS · R01 GM068101 · United States
NIGMS NIH HHS · R01 GM068101-01 · United States
External Links
PubMed source DOI: 10.1091/mbc.e04-09-0787 PMC Full Text View journal details
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