The DHC1b (DHC2) isoform of cytoplasmic dynein is required for flagellar assembly.
Dyneins are microtubule-based molecular motors involved in many different types of cell movement. Most dynein heavy chains (DHCs) clearly group into cytoplasmic or axonemal isoforms. However, DHC1b has been enigmatic. To learn more about this isoform, we isolated Chlamydomonas cDNA clones encoding a portion of DHC1b, and used these clones to identify a Chlamydomonas cell line with a deletion mutation in DHC1b. The mutant grows normally and appears to have a normal Golgi apparatus, but has very short flagella. The deletion also results in a massive redistribution of raft subunits from a peri-basal body pool (Cole, D.G., D.R. Diener, A.L. Himelblau, P.L. Beech, J.C. Fuster, and J.L. Rosenbaum. 1998. J. Cell Biol. 141:993-1008) to the flagella. Rafts are particles that normally move up and down the flagella in a process known as intraflagellar transport (IFT) (Kozminski, K.G., K.A. Johnson, P. Forscher, and J.L. Rosenbaum. 1993. Proc. Natl. Acad. Sci. USA. 90:5519-5523), which is essential for assembly and maintenance of flagella. The redistribution of raft subunits apparently occurs due to a defect in the retrograde component of IFT, suggesting that DHC1b is the motor for retrograde IFT. Consistent with this, Western blots indicate that DHC1b is present in the flagellum, predominantly in the detergent- and ATP-soluble fractions. These results indicate that DHC1b is a cytoplasmic dynein essential for flagellar assembly, probably because it is the motor for retrograde IFT.
Pazour
G J
GJ
Department of Cell Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
Dickert
B L
BL
Witman
G B
GB
D000595:Amino Acid Sequence; D000818:Animals; D001693:Biological Transport, Active; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D056445:Cytoplasmic Dyneins; D004398:Dyneins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism* / Q000648:ultrastructure; D017353:Gene Deletion; D017125:Genes, Protozoan; D006056:Golgi Apparatus / Q000648:ultrastructure; D008854:Microscopy, Electron; D020409:Molecular Motor Proteins; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid