The Vfl1 Protein in Chlamydomonas localizes in a rotationally asymmetric pattern at the distal ends of the basal bodies
| Autor: | Mark A. Sanders, Paul A. Lefebvre, Carolyn D. Silflow, Lai Wa Tam, Susan Tousey, Mark Borodovsky, Wei Chien Wu, Matthew J. LaVoie |
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| Rok vydání: | 2001 |
| Předmět: |
Centriole
Molecular Sequence Data Protozoan Proteins Flagellum basal body 03 medical and health sciences 0302 clinical medicine Microtubule Basal body Animals centriole Amino Acid Sequence Gene Alleles 030304 developmental biology 0303 health sciences biology Cilium Chlamydomonas Algal Proteins cilia Cell Biology Immunogold labelling Anatomy biology.organism_classification Cell biology Original Article flagella 030217 neurology & neurosurgery |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 0021-9525 |
| Popis: | 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 (Hoops, H.J., and G.B. Witman. 1983. J. Cell Biol. 97:902–908). The vfl1 mutation results in variable numbers and positioning of flagella and basal bodies (Adams, G.M.W., R.L. Wright, and J.W. Jarvik. 1985. J. Cell Biol. 100:955–964). 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. |
| Databáze: | OpenAIRE |
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