Mutations in alpha-tubulin promote basal body maturation and flagellar assembly in the absence of delta-tubulin.
We have isolated suppressors of the deletion allele of delta-tubulin, uni3-1, in the biflagellate green alga Chlamydomonas reinhardtii. The deletion of delta-tubulin produces cells that assemble zero, one or two flagella and have basal bodies composed primarily of doublet rather than triplet microtubules. Flagellar number is completely restored in the suppressed strains. Most of the uni3-1 suppressors map to the TUA2 locus, which encodes alpha2-tubulin. Twelve independent tua2 mutations were sequenced. Amino acids D205 or A208, which are nearly invariant residues in alpha-tubulin, were altered. The tua2 mutations on their own have a second phenotype - they make the cells colchicine supersensitive. Colchicine supersensitivity itself is not needed for suppression and colchicine cannot phenocopy the suppression. The suppressors partially restore the assembly of triplet microtubules. These results suggest that the delta-tubulin plays two roles: it is needed for extension or stability of the triplet microtubule and also for early maturation of basal bodies. We suggest that the mutant alpha-tubulin promotes the early maturation of the basal body in the absence of delta-tubulin, perhaps through interactions with other partners, and this allows assembly of the flagella.
Fromherz
Sylvia
S
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Boulder, CO 80309-0347, USA.
Giddings
Thomas H
TH
Gomez-Ospina
Natalia
N
Dutcher
Susan K
SK
D000483:Alleles; D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism; D003078:Colchicine / Q000494:pharmacology; D018390:Gene Targeting; D008854:Microscopy, Electron; D008868:Microtubule Proteins / Q000187:drug effects / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D008961:Models, Structural; D009154:Mutation; D010641:Phenotype; D014404:Tubulin / Q000235:genetics / Q000378:metabolism*
D008868:Microtubule Proteins; D014404:Tubulin; D003078:Colchicine