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Published online 2 April 2001. doi:10.1083/jcb.153.1.63
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© The Rockefeller University Press, 0021-9525/2001//63 $5.00
The Journal of Cell Biology, Volume 153, Number 1, , 2001 63-74

Original Article

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



Carolyn D. Silflowa,b, Matthew LaVoiea, Lai-Wa Tama, Susan Touseya, Mark Sandersa, Wei-chien Wua, Mark Borodovskyc, and Paul A. Lefebvrea,b

a Department of Genetics, Cell Biology and Development
b Department of Plant Biology, University of Minnesota, St. Paul, Minnesota 55108
c School of Biology, Georgia Institute of Technology, Atlanta, Georgia 30332
University of Minnesota, 250 Bioscience Center, 1445 Gortner Ave., St. Paul, MN 55108.(612) 625-5754(612) 624-0729

carolyn{at}biosci.cbs.umn.edu

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 {alpha}-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.

Key Words: basal body • flagella • cilia • centriole • Chlamydomonas

© 2001 The Rockefeller University Press

Abbreviations used in this paper: DIC, differential interference contrast; HA, hemagglutinin; LRR, leucine-rich repeat; NFAp, nucleoflagellar apparatus; RT, reverse transcriptase.


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