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PMID: 10806107 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Fa1p is a 171 kDa protein essential for axonemal microtubule severing in Chlamydomonas.

Journal of cell science ·Vol. 113 ( Pt 11) ·2000-06-00 ·Pages 1963-71

Finst RJ, Kim PJ, Griffis ER, Quarmby LM

Abstract

A key event in deflagellation or deciliation is the severing of the nine outer-doublet axonemal microtubules at a specific site in the flagellar transition zone. Previous genetic analysis revealed three genes that are essential for deflagellation in Chlamydomonas. We have now identified the first of these products, Fa1p, a protein required for Ca(2+)-dependent, axonemal microtubule severing. Genetic mapping and the availability of a tagged allele allowed us to physically map the gene to the centromere-proximal domain of the mating-type locus. We identified clones of Chlamydomonas genomic DNA that rescued the Ca(2+)-dependent axonemal microtubule severing defect of fa1 mutants. The FA1 cDNA, obtained by RT-PCR, encodes a novel protein of 171 kDa, which is predicted to contain an amino-terminal coiled-coil domain and three Ca(2+)/calmodulin binding domains. By western analysis and subcellular fractionation, the FA1 product is enriched in flagellar-basal body complexes. Based on these observations and previous studies, we hypothesize that a Ca(2+)-activated, Ca(2+)-binding protein binds Fa1p leading ultimately to the activation of axonemal microtubule severing.

MeSH Terms
Amino Acid Sequence Animals Bacterial Proteins/genetics,metabolism Blotting, Northern Calcium/metabolism Calcium Signaling/physiology Calmodulin/metabolism Chlamydomonas reinhardtii/genetics,metabolism,ultrastructure Chromosome Mapping Cloning, Molecular DNA, Complementary/analysis Flagella/physiology Microtubule Proteins/genetics,metabolism Microtubules/physiology Molecular Sequence Data Molecular Weight Mutagenesis/physiology Protein Structure, Secondary RNA, Messenger/analysis Sequence Analysis, DNA
Chemicals
Bacterial Proteins Calmodulin DNA, Complementary Fa1 protein, Clamydomonas Microtubule Proteins RNA, Messenger Calcium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Finst R J
Department of Cell Biology and Graduate Program in Biochemistry, Cell and Developmental Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Kim P J
Griffis E R
Quarmby L M
Article Info
Journal
Journal of cell science
Abbr.
J Cell Sci
ISSN
0021-9533
Published
2000-06-00
Pages
1963-71
Language
English
Region
England
NLM ID
0052457
Subset
IM
Grants
NIGMS NIH HHS · T32 GM-08367 · United States
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