Oda5p, a novel axonemal protein required for assembly of the outer dynein arm and an associated adenylate kinase.

Abstract:

:Of the uncloned ODA genes required for outer dynein arm assembly in Chlamydomonas, ODA5 and ODA10 are of particular interest because they do not encode known subunits of the outer arm or the outer dynein arm-docking complex (ODA-DC), and because genetic studies suggest their products interact. Beginning with a tagged oda5 allele, we isolated genomic and cDNA clones of the wild-type gene. ODA5 predicts a novel, 66-kDa coiled-coil protein. Immunoblotting indicates Oda5p is an axonemal component that assembles onto the axoneme independently of the outer arm and ODA-DC and is uniquely missing in oda5 and oda10 axonemes. Oda5p is released from the axoneme by extraction with 0.6 M KCl, but the soluble Oda5p does not cosediment with the outer dynein arm/ODA-DC in sucrose gradients. Quantitative mass spectrometry by using isotope coded affinity tagging revealed that a previously unidentified adenylate kinase is reduced 35-50% in oda5 flagella. Direct enzymatic assays demonstrated a comparable reduction in adenylate kinase activity in oda5 flagella, and also in oda10 flagella, but not in flagella of other oda mutants. We propose that Oda5p is part of a novel axonemal complex that is required for outer arm assembly and anchors adenylate kinase in proximity to the arm.

journal_name

Mol Biol Cell

authors

Wirschell M,Pazour G,Yoda A,Hirono M,Kamiya R,Witman GB

doi

10.1091/mbc.e03-11-0820

keywords:

subject

Has Abstract

pub_date

2004-06-01 00:00:00

pages

2729-41

issue

6

eissn

1059-1524

issn

1939-4586

pii

E03-11-0820

journal_volume

15

pub_type

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