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The Chlamydomonas reinhardtii ODA3 gene encodes a protein of the outer dynein arm docking complex.

Koutoulis A, Pazour GJ, Wilkerson CG, Inaba K, Sheng H, Takada S, Witman GB - J. Cell Biol. (1997)

Bottom Line: Kamiya. 1994.The cDNA sequence predicts a novel 83. 4-kD protein with extensive coiled-coil domains.The ODA-DC contains three polypeptides; direct amino acid sequencing indicates that the largest of these polypeptides corresponds to ODA3.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Science, The University of Tasmania, Hobart TAS 7001 Australia.

ABSTRACT
We have used an insertional mutagenesis/ gene tagging technique to generate new Chlamydomonas reinhardtii mutants that are defective in assembly of the uter ynein rm. Among 39 insertional oda mutants characterized, two are alleles of the previously uncloned ODA3 gene, one is an allele of the uncloned ODA10 gene, and one represents a novel ODA gene (termed ODA12). ODA3 is of particular interest because it is essential for assembly of both the outer dynein arm and the outer dynein arm docking complex (ODA-DC) onto flagellar doublet microtubules (Takada, S., and R. Kamiya. 1994. J. Cell Biol. 126:737- 745). Beginning with the inserted DNA as a tag, the ODA3 gene and a full-length cDNA were cloned. The cloned gene rescues the phenotype of oda3 mutants. The cDNA sequence predicts a novel 83. 4-kD protein with extensive coiled-coil domains. The ODA-DC contains three polypeptides; direct amino acid sequencing indicates that the largest of these polypeptides corresponds to ODA3. This protein is likely to have an important role in the precise positioning of the outer dynein arms on the flagellar axoneme.

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Nucleic acid and deduced amino acid sequence of  ODA3. The numbers on the lefthand and righthand sides refer to  nucleotides and amino acid residues, respectively. In-frame stop  codons before and after the open reading frame are marked with  asterisks. Underlined amino acids correspond to sequence obtained directly from tryptic fragments of the ∼105-kD polypeptide of the ODA-DC. Dotted underlines mark the imperfect 11– amino acid tandem repeat. These sequence data are available from  GenBank/EMBL/DDBJ under accession number AF001309.
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Figure 5: Nucleic acid and deduced amino acid sequence of ODA3. The numbers on the lefthand and righthand sides refer to nucleotides and amino acid residues, respectively. In-frame stop codons before and after the open reading frame are marked with asterisks. Underlined amino acids correspond to sequence obtained directly from tryptic fragments of the ∼105-kD polypeptide of the ODA-DC. Dotted underlines mark the imperfect 11– amino acid tandem repeat. These sequence data are available from GenBank/EMBL/DDBJ under accession number AF001309.

Mentions: The ∼3.0-kb clone was sequenced in both directions and found to contain a 1,797-bp open reading frame. The putative initiation codon was preceded by two in-frame stop codons located 45 and 99 nucleotides upstream (Fig. 5). The open reading frame ended with an amber stop codon (TAG). However, it was noted that an additional in-frame 447-bp open reading frame was present immediately after the amber stop codon. Both open reading frames exhibited the strong codon bias typical of C. reinhardtii coding sequence (Williams et al., 1989; Mitchell and Brown, 1994). To understand this phenomenon, the ∼2.4-kb clone was sequenced through this region. The sequence was identical in both clones except that the TAG codon was CAG in the ∼2.4-kb clone. We conclude that the amber stop codon in the ∼3.0-kb clone was the result of a point mutation that occurred during cDNA library construction. The correct codon should be CAG, resulting in a 2,247-bp open reading frame terminating in TAA (Fig. 5). Two additional inframe stop codons (TAA and TGA) were located 24 and 66 nucleotides downstream from the first TAA, providing further evidence that the first TAA represents the actual termination codon.


The Chlamydomonas reinhardtii ODA3 gene encodes a protein of the outer dynein arm docking complex.

Koutoulis A, Pazour GJ, Wilkerson CG, Inaba K, Sheng H, Takada S, Witman GB - J. Cell Biol. (1997)

Nucleic acid and deduced amino acid sequence of  ODA3. The numbers on the lefthand and righthand sides refer to  nucleotides and amino acid residues, respectively. In-frame stop  codons before and after the open reading frame are marked with  asterisks. Underlined amino acids correspond to sequence obtained directly from tryptic fragments of the ∼105-kD polypeptide of the ODA-DC. Dotted underlines mark the imperfect 11– amino acid tandem repeat. These sequence data are available from  GenBank/EMBL/DDBJ under accession number AF001309.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2136212&req=5

Figure 5: Nucleic acid and deduced amino acid sequence of ODA3. The numbers on the lefthand and righthand sides refer to nucleotides and amino acid residues, respectively. In-frame stop codons before and after the open reading frame are marked with asterisks. Underlined amino acids correspond to sequence obtained directly from tryptic fragments of the ∼105-kD polypeptide of the ODA-DC. Dotted underlines mark the imperfect 11– amino acid tandem repeat. These sequence data are available from GenBank/EMBL/DDBJ under accession number AF001309.
Mentions: The ∼3.0-kb clone was sequenced in both directions and found to contain a 1,797-bp open reading frame. The putative initiation codon was preceded by two in-frame stop codons located 45 and 99 nucleotides upstream (Fig. 5). The open reading frame ended with an amber stop codon (TAG). However, it was noted that an additional in-frame 447-bp open reading frame was present immediately after the amber stop codon. Both open reading frames exhibited the strong codon bias typical of C. reinhardtii coding sequence (Williams et al., 1989; Mitchell and Brown, 1994). To understand this phenomenon, the ∼2.4-kb clone was sequenced through this region. The sequence was identical in both clones except that the TAG codon was CAG in the ∼2.4-kb clone. We conclude that the amber stop codon in the ∼3.0-kb clone was the result of a point mutation that occurred during cDNA library construction. The correct codon should be CAG, resulting in a 2,247-bp open reading frame terminating in TAA (Fig. 5). Two additional inframe stop codons (TAA and TGA) were located 24 and 66 nucleotides downstream from the first TAA, providing further evidence that the first TAA represents the actual termination codon.

Bottom Line: Kamiya. 1994.The cDNA sequence predicts a novel 83. 4-kD protein with extensive coiled-coil domains.The ODA-DC contains three polypeptides; direct amino acid sequencing indicates that the largest of these polypeptides corresponds to ODA3.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Science, The University of Tasmania, Hobart TAS 7001 Australia.

ABSTRACT
We have used an insertional mutagenesis/ gene tagging technique to generate new Chlamydomonas reinhardtii mutants that are defective in assembly of the uter ynein rm. Among 39 insertional oda mutants characterized, two are alleles of the previously uncloned ODA3 gene, one is an allele of the uncloned ODA10 gene, and one represents a novel ODA gene (termed ODA12). ODA3 is of particular interest because it is essential for assembly of both the outer dynein arm and the outer dynein arm docking complex (ODA-DC) onto flagellar doublet microtubules (Takada, S., and R. Kamiya. 1994. J. Cell Biol. 126:737- 745). Beginning with the inserted DNA as a tag, the ODA3 gene and a full-length cDNA were cloned. The cloned gene rescues the phenotype of oda3 mutants. The cDNA sequence predicts a novel 83. 4-kD protein with extensive coiled-coil domains. The ODA-DC contains three polypeptides; direct amino acid sequencing indicates that the largest of these polypeptides corresponds to ODA3. This protein is likely to have an important role in the precise positioning of the outer dynein arms on the flagellar axoneme.

Show MeSH
Related in: MedlinePlus