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A subunit of the dynein regulatory complex in Chlamydomonas is a homologue of a growth arrest-specific gene product.

Rupp G, Porter ME - J. Cell Biol. (2003)

Bottom Line: The pf2-4 mutant displays an altered waveform that results in slow swimming cells.PF2 is a coiled-coil protein that shares significant homology with a mammalian growth arrest-specific gene product (Gas11/Gas8) and a trypanosome protein known as trypanin.The expression of Gas8/Gas11 transcripts in a wide range of tissues may also indicate a potential role for PF2-related proteins in other microtubule-based structures.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota Medical School, 6-160 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455, USA.

ABSTRACT
The dynein regulatory complex (DRC) is an important intermediate in the pathway that regulates flagellar motility. To identify subunits of the DRC, we characterized a Chlamydomonas motility mutant obtained by insertional mutagenesis. The pf2-4 mutant displays an altered waveform that results in slow swimming cells. EM analysis reveals defects in DRC structure that can be rescued by reintroduction of the wild-type PF2 gene. Immunolocalization studies show that the PF2 protein is distributed along the length of the axoneme, where it is part of a discrete complex of polypeptides. PF2 is a coiled-coil protein that shares significant homology with a mammalian growth arrest-specific gene product (Gas11/Gas8) and a trypanosome protein known as trypanin. PF2 and its homologues appear to be universal components of motile axonemes that are required for DRC assembly and the regulation of flagellar motility. The expression of Gas8/Gas11 transcripts in a wide range of tissues may also indicate a potential role for PF2-related proteins in other microtubule-based structures.

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Localization of the epitope-tagged PF2 protein in Chlamydomonas flagella. The epitope-tagged, rescued strain, pf2-4r:3HA, was stained with an antibody against the HA tag and then imaged by differential interference contrast (A and C) or indirect immunofluorescence light microscopy (B and D). The HA-tagged PF2 is present along the entire length of the flagella in pf2-4r:3HA cells and can also been seen in two spots in the basal body region (B and D). pf2-4 mutant (E and F) or wild-type cells (G and H) labeled with the HA antibody or secondary antibody alone (not depicted) show only background cell body autofluorescence.
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fig5: Localization of the epitope-tagged PF2 protein in Chlamydomonas flagella. The epitope-tagged, rescued strain, pf2-4r:3HA, was stained with an antibody against the HA tag and then imaged by differential interference contrast (A and C) or indirect immunofluorescence light microscopy (B and D). The HA-tagged PF2 is present along the entire length of the flagella in pf2-4r:3HA cells and can also been seen in two spots in the basal body region (B and D). pf2-4 mutant (E and F) or wild-type cells (G and H) labeled with the HA antibody or secondary antibody alone (not depicted) show only background cell body autofluorescence.

Mentions: Mutant and rescued cells were stained with antibodies against the HA epitope to analyze the subcellular distribution of PF2. Analysis of pf2 mutant and pf2 rescued cells by differential interference contrast microscopy demonstrated that both strains assemble full-length flagella (Fig. 5, A, C, and E)Figure 5.


A subunit of the dynein regulatory complex in Chlamydomonas is a homologue of a growth arrest-specific gene product.

Rupp G, Porter ME - J. Cell Biol. (2003)

Localization of the epitope-tagged PF2 protein in Chlamydomonas flagella. The epitope-tagged, rescued strain, pf2-4r:3HA, was stained with an antibody against the HA tag and then imaged by differential interference contrast (A and C) or indirect immunofluorescence light microscopy (B and D). The HA-tagged PF2 is present along the entire length of the flagella in pf2-4r:3HA cells and can also been seen in two spots in the basal body region (B and D). pf2-4 mutant (E and F) or wild-type cells (G and H) labeled with the HA antibody or secondary antibody alone (not depicted) show only background cell body autofluorescence.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172716&req=5

fig5: Localization of the epitope-tagged PF2 protein in Chlamydomonas flagella. The epitope-tagged, rescued strain, pf2-4r:3HA, was stained with an antibody against the HA tag and then imaged by differential interference contrast (A and C) or indirect immunofluorescence light microscopy (B and D). The HA-tagged PF2 is present along the entire length of the flagella in pf2-4r:3HA cells and can also been seen in two spots in the basal body region (B and D). pf2-4 mutant (E and F) or wild-type cells (G and H) labeled with the HA antibody or secondary antibody alone (not depicted) show only background cell body autofluorescence.
Mentions: Mutant and rescued cells were stained with antibodies against the HA epitope to analyze the subcellular distribution of PF2. Analysis of pf2 mutant and pf2 rescued cells by differential interference contrast microscopy demonstrated that both strains assemble full-length flagella (Fig. 5, A, C, and E)Figure 5.

Bottom Line: The pf2-4 mutant displays an altered waveform that results in slow swimming cells.PF2 is a coiled-coil protein that shares significant homology with a mammalian growth arrest-specific gene product (Gas11/Gas8) and a trypanosome protein known as trypanin.The expression of Gas8/Gas11 transcripts in a wide range of tissues may also indicate a potential role for PF2-related proteins in other microtubule-based structures.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota Medical School, 6-160 Jackson Hall, 321 Church Street SE, Minneapolis, MN 55455, USA.

ABSTRACT
The dynein regulatory complex (DRC) is an important intermediate in the pathway that regulates flagellar motility. To identify subunits of the DRC, we characterized a Chlamydomonas motility mutant obtained by insertional mutagenesis. The pf2-4 mutant displays an altered waveform that results in slow swimming cells. EM analysis reveals defects in DRC structure that can be rescued by reintroduction of the wild-type PF2 gene. Immunolocalization studies show that the PF2 protein is distributed along the length of the axoneme, where it is part of a discrete complex of polypeptides. PF2 is a coiled-coil protein that shares significant homology with a mammalian growth arrest-specific gene product (Gas11/Gas8) and a trypanosome protein known as trypanin. PF2 and its homologues appear to be universal components of motile axonemes that are required for DRC assembly and the regulation of flagellar motility. The expression of Gas8/Gas11 transcripts in a wide range of tissues may also indicate a potential role for PF2-related proteins in other microtubule-based structures.

Show MeSH
Related in: MedlinePlus