The Vfl1 Protein in Chlamydomonas Localizes in a Rotationally Asymmetric Pattern at the Distal Ends of the Basal Bodies

In the unicellular alga Chlamydomonas, two anterior flagella are positioned with 180° rotational symmetry, such that the flagella beat with the effective strokes in opposite directions. The vfl1 mutation results in variable numbers and positioning of flagella and basal bodies. Using a tagged allele,...

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Bibliographic Details
Published in:The Journal of cell biology 2001-04, Vol.153 (1), p.63-74
Main Authors: Silflow, Carolyn D., LaVoie, Matthew, Tam, Lai-Wa, Tousey, Susan, Sanders, Mark, Wu, Wei-chien, Borodovsky, Mark, Lefebvre, Paul A.
Format: Article
Language:English
Subjects:
Algal Proteins - analysis
Algal Proteins - genetics
Alleles
Amino Acid Sequence
Animals
Antibodies
Cell cycle
Cell lines
Cells
Cellular biology
Chlamydomonas
Chlamydomonas - chemistry
Chlamydomonas - genetics
DNA
Flagella
Freshwater
Genetic mutation
Microtubules
Molecular Sequence Data
Neurotransmitters
Original
Phenotypes
Proteins
Protozoan Proteins - analysis
Protozoan Proteins - genetics
Rotating bodies
Vfl1 protein
VLF1 gene
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Summary:In the unicellular alga Chlamydomonas, two anterior flagella are positioned with 180° rotational symmetry, such that the flagella beat with the effective strokes in opposite directions. The vfl1 mutation results in variable numbers and positioning of flagella and basal bodies. Using a tagged allele, we cloned the VFL1 gene that encodes a protein of 128 kD with five leucine-rich repeat sequences near the NH2 terminus and a large α-helical-coiled coil domain at the COOH terminus. An epitope-tagged gene construct rescued the mutant phenotype and expressed a tagged protein (Vfl1p) that copurified with basal body flagellar apparatuses. Immunofluorescence experiments showed that Vfl1p localized with basal bodies and probasal bodies. Immunogold labeling localized Vfl1p inside the lumen of the basal body at the distal end. Distribution of gold particles was rotationally asymmetric, with most particles located near the doublet microtubules that face the opposite basal body. The mutant phenotype, together with the localization results, suggest that Vfl1p plays a role in establishing the correct rotational orientation of basal bodies. Vfl1p is the first reported molecular marker of the rotational asymmetry inherent to basal bodies.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.153.1.63