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Published 10 November 2003. doi:10.1083/jcb.200307143
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© The Rockefeller University Press, 0021-9525/2003/11/597 $8.00
The Journal of Cell Biology, Volume 163, Number 3, 597-607

Article

 Defective flagellar assembly and length regulation in LF3 null mutants in Chlamydomonas

Lai-Wa Tam1, William L. Dentler2 and Paul A. Lefebvre1

1 Department of Plant Biology, University of Minnesota, St. Paul, MN 55108
2 Department of Molecular Biosciences, University of Kansas, Lawrence, KS 66045

Address correspondence to Paul A. Lefebvre, Dept. of Plant Biology, University of Minnesota, 250 Biological Sciences Center, 1445 Gortner Ave., St. Paul, MN 55108. Tel.: (612) 624-0729. Fax: (612) 625-1738. email: pete{at}biosci.cbs.umn.edu

Four long-flagella (LF) genes are important for flagellar length control in Chlamydomonas reinhardtii. Here, we characterize two new null lf3 mutants whose phenotypes are different from previously identified lf3 mutants. These null mutants have unequal-length flagella that assemble more slowly than wild-type flagella, though their flagella can also reach abnormally long lengths. Prominent bulges are found at the distal ends of short, long, and regenerating flagella of these mutants. Analysis of the flagella by electron and immunofluorescence microscopy and by Western blots revealed that the bulges contain intraflagellar transport complexes, a defect reported previously (for review see Cole, D.G., 2003. Traffic. 4:435–442) in a subset of mutants defective in intraflagellar transport. We have cloned the wild-type LF3 gene and characterized a hypomorphic mutant allele of LF3. LF3p is a novel protein located predominantly in the cell body. It cosediments with the product of the LF1 gene in sucrose density gradients, indicating that these proteins may form a functional complex to regulate flagellar length and assembly.

Key Words: cilia; intraflagellar transport; organelle size control; unequal-length flagella

Abbreviations used in this paper: DIC, differential interference contrast; IFT, intraflagellar transport; Ulf, unequal-length flagella; WT, wild-type.


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