The Chlamydomonas MBO2 locus encodes a conserved coiled-coil protein important for flagellar waveform conversion.
Chlamydomonas flagella can undergo a calcium-dependent conversion between an asymmetric ciliary waveform and a symmetric flagellar waveform. Mutations at three MBO loci abolish the predominant ciliary waveform and result in cells that move backward only with the flagellar waveform. We have cloned and characterized the MBO2 gene. It encodes a novel protein with extensive alpha-helical coiled-coils and two leucine zippers. Sequences highly similar to MBO2p were found in a variety of organisms with cilia and flagella, suggesting that the MBO2 gene function may be conserved in many diverse taxa. Antibodies to MBO2p recognized an axonemal protein of 110 kDa, which appeared to be tightly associated with doublet microtubules. The protein was present in flagella of a variety of paralyzed flagellar mutants that lacked different axonemal structures, indicating that MBO2p is a component of a previously uncharacterized flagellar protein complex. In contrast to the earlier suggestion that the MBO2 gene may encode a component of an intramicrotubular beak-like structure present only proximally in flagella, we localized an epitope-tagged MBO2p along the entire length of the flagella. Moreover, the insertion of a hemagglutinin (HA) epitope in the conserved C-terminal domain of MBO2p reduced the swimming velocity of cells transformed with the epitope-tagged gene. These results indicate that MBO2p may play a role both in the assembly of the beak-like structure and the regulation of the force-generation machinery during the ciliary beat.
Tam
Lai-Wa
LW
Department of Genetics, Cell Biology and Development, University of Minnesota, St. Paul 55108, USA.
Lefebvre
Paul A
PA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002696:Chlamydomonas / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D016054:DNA, Protozoan; D005407:Flagella / Q000502:physiology* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting; D008868:Microtubule Proteins / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger; D016053:RNA, Protozoan; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D016054:DNA, Protozoan; D008868:Microtubule Proteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D002118:Calcium