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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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Structural defects in pf2 axonemes. Longitudinal images of wild-type and mutant axonemes were aligned and computer image averaged. Shown here are grand averages of axonemes isolated from (A) wild type, (B) pf2-4, (C), pf2-1, and (D) a rescued pf2 strain (pf2-4r). The arrows in A–D indicate the region of the 96-nm repeat that is occupied by the DRC structure. (E) Model of axoneme structures within the 96-nm axoneme repeat. The proximal and distal RSs are labeled S1 and S2, respectively; the outer dynein arms (OA) are shown on top, and the DRC is indicated as a crescent-shaped structure above the distal RS (S2). Difference plots between wild type (wt) and pf2-4 (F), and wt and pf2-1 (G) demonstrate that the DRC is missing in both mutants. Note that the DRC is restored in the rescued strain, pf2-4r (D).
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fig3: Structural defects in pf2 axonemes. Longitudinal images of wild-type and mutant axonemes were aligned and computer image averaged. Shown here are grand averages of axonemes isolated from (A) wild type, (B) pf2-4, (C), pf2-1, and (D) a rescued pf2 strain (pf2-4r). The arrows in A–D indicate the region of the 96-nm repeat that is occupied by the DRC structure. (E) Model of axoneme structures within the 96-nm axoneme repeat. The proximal and distal RSs are labeled S1 and S2, respectively; the outer dynein arms (OA) are shown on top, and the DRC is indicated as a crescent-shaped structure above the distal RS (S2). Difference plots between wild type (wt) and pf2-4 (F), and wt and pf2-1 (G) demonstrate that the DRC is missing in both mutants. Note that the DRC is restored in the rescued strain, pf2-4r (D).

Mentions: To determine if the pf2-4 motility phenotype was also associated with structural defects in the DRC, axonemes were isolated from wild-type and pf2-4 cells and prepared for EM. Analysis of longitudinal sections with image-averaging procedures revealed a defect in a crescent-shaped structure located close to the second RS, within the 96-nm axoneme repeat (Fig. 3, A and B)Figure 3.


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)

Structural defects in pf2 axonemes. Longitudinal images of wild-type and mutant axonemes were aligned and computer image averaged. Shown here are grand averages of axonemes isolated from (A) wild type, (B) pf2-4, (C), pf2-1, and (D) a rescued pf2 strain (pf2-4r). The arrows in A–D indicate the region of the 96-nm repeat that is occupied by the DRC structure. (E) Model of axoneme structures within the 96-nm axoneme repeat. The proximal and distal RSs are labeled S1 and S2, respectively; the outer dynein arms (OA) are shown on top, and the DRC is indicated as a crescent-shaped structure above the distal RS (S2). Difference plots between wild type (wt) and pf2-4 (F), and wt and pf2-1 (G) demonstrate that the DRC is missing in both mutants. Note that the DRC is restored in the rescued strain, pf2-4r (D).
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Related In: Results  -  Collection

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

fig3: Structural defects in pf2 axonemes. Longitudinal images of wild-type and mutant axonemes were aligned and computer image averaged. Shown here are grand averages of axonemes isolated from (A) wild type, (B) pf2-4, (C), pf2-1, and (D) a rescued pf2 strain (pf2-4r). The arrows in A–D indicate the region of the 96-nm repeat that is occupied by the DRC structure. (E) Model of axoneme structures within the 96-nm axoneme repeat. The proximal and distal RSs are labeled S1 and S2, respectively; the outer dynein arms (OA) are shown on top, and the DRC is indicated as a crescent-shaped structure above the distal RS (S2). Difference plots between wild type (wt) and pf2-4 (F), and wt and pf2-1 (G) demonstrate that the DRC is missing in both mutants. Note that the DRC is restored in the rescued strain, pf2-4r (D).
Mentions: To determine if the pf2-4 motility phenotype was also associated with structural defects in the DRC, axonemes were isolated from wild-type and pf2-4 cells and prepared for EM. Analysis of longitudinal sections with image-averaging procedures revealed a defect in a crescent-shaped structure located close to the second RS, within the 96-nm axoneme repeat (Fig. 3, A and B)Figure 3.

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