A novel dynein light intermediate chain colocalizes with the retrograde motor for intraflagellar transport at sites of axoneme assembly in Chlamydomonas and mammalian cells

Catherine A. Perrone, Douglas Tritschler, Patrick Taulman, Raqual Bower, Bradley K. Yoder, Mary E. Porter

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

The assembly of cilia and flagella depends on bidirectional intraflagellar transport (IFT). Anterograde IFT is driven by kinesin II, whereas retrograde IFT requires cytoplasmic dynein 1b (cDHC1b). Little is known about how cDHC1b interacts with its cargoes or how it is regulated. Recent work identified a novel dynein light intermediate chain (D2LIC) that colocalized with the mammalian cDHClb homolog DHC2 in the centrosomal region of cultured cells. To see whether the LIC might play a role in IFT, we characterized the gene encoding the Chlamydomonas homolog of D2LIC and found its expression is up-regulated in response to deflagellation. We show that the LIC subunit copurifies with cDHC1b during flagellar isolation, dynein extraction, sucrose density centrifugation, and immunoprecipitation. Immunocytochemistry reveals that the LIC colocalizes with cDHC1b in the basal body region and along the length of flagella in wild-type cells. Localization of the complex is altered in a collection of retrograde IFT and length control mutants, which suggests that the affected gene products directly or indirectly regulate cDHC1b activity. The mammalian DHC2 and D2LIC also colocalize in the apical cytoplasm and axonemes of ciliated epithelia in the lung, brain, and efferent duct. These studies, together with the identification of an LIC mutation, xbx-1(ok279), which disrupts retrograde IFT in Caenorhabditis elegans, indicate that the novel LIC is a component of the cDHClb/DHC2 retrograde IFT motor in a variety of organisms.

Original languageEnglish (US)
Pages (from-to)2041-2056
Number of pages16
JournalMolecular biology of the cell
Volume14
Issue number5
DOIs
StatePublished - May 1 2003

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Axoneme
Chlamydomonas
Dyneins
Flagella
Cytoplasmic Dyneins
Basal Bodies
Body Regions
Cilia
Caenorhabditis elegans
Centrifugation
Immunoprecipitation
Genes
Sucrose
Cultured Cells
Cytoplasm
Epithelium
Immunohistochemistry
Lung
Mutation
Brain

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A novel dynein light intermediate chain colocalizes with the retrograde motor for intraflagellar transport at sites of axoneme assembly in Chlamydomonas and mammalian cells. / Perrone, Catherine A.; Tritschler, Douglas; Taulman, Patrick; Bower, Raqual; Yoder, Bradley K.; Porter, Mary E.

In: Molecular biology of the cell, Vol. 14, No. 5, 01.05.2003, p. 2041-2056.

Research output: Contribution to journalArticle

Perrone, Catherine A. ; Tritschler, Douglas ; Taulman, Patrick ; Bower, Raqual ; Yoder, Bradley K. ; Porter, Mary E. / A novel dynein light intermediate chain colocalizes with the retrograde motor for intraflagellar transport at sites of axoneme assembly in Chlamydomonas and mammalian cells. In: Molecular biology of the cell. 2003 ; Vol. 14, No. 5. pp. 2041-2056.
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