The two alpha-tubulin genes of Chlamydomonas reinhardi code for slightly different proteins.

Silflow, Carolyn D.; Chisholm, Rex L.; Conner, Timothy W.; Ranum, Laura P.W.

doi:10.1128/mcb.5.9.2389

Cited by 174 publications

(88 citation statements)

References 44 publications

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“…The motif TGTAA, considered to be a potential polyadenylation signal in C. reinhardtii (Silflow et al 1985), was found 18 bases upstream from the 3'-end and also at position 1,225 in the cDNA sequence.…”

Section: Isolation and Sequence Analysis Of The Aox1 Cdnamentioning

confidence: 99%

Characterization of two genes encoding the mitochondrial alternative oxidase in Chlamydomonas reinhardtii

Dinant¹,

Baurain²,

Coosemans³

et al. 2001

Current Genetics

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Two cDNA clones (AOX1 and AOX2) and the corresponding genes encoding the alternative oxidases (AOXs) from Chlamydomonas reinhardtii were isolated and sequenced. The cDNAs, AOX1 and AOX2, contained open reading frames (ORFs) encoding putative proteins of 360 amino acids and 347 amino acids, respectively. For each of the ORFs, a potential mitochondrial-targeting sequence was found in the 5'-end regions. In comparison to AOX enzymes from plants and fungi, the predicted amino acid sequences of the ORFs showed their highest degree of identity with proteins from Aspergillus niger (38.1% and 37.2%) and Ajellomyces capsulatus (37% and 34.9%). Several residues supposed either to be Fe ligands or to be involved in the ubiquinol-binding site were fully conserved in both C. reinhardtii putative AOX proteins. In contrast, a cysteine residue conserved in the sequences of all higher plants and probably involved in the regulation of the enzyme activity was missing both from the AOX1 and AOX2 amino acid sequences and from protein sequences from various other microorganisms. The transcriptional expression of the AOX1 and AOX2 genes in wild-type cells and in mutant cells deficient in mitochondrial complex III activity was also investigated.

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Section: Isolation and Sequence Analysis Of The Aox1 Cdnamentioning

confidence: 99%

Characterization of two genes encoding the mitochondrial alternative oxidase in Chlamydomonas reinhardtii

Dinant¹,

Baurain²,

Coosemans³

et al. 2001

Current Genetics

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“…The 5Ј end of the RNA transcript extends at least 272 base pairs upstream of the putative start codon. A putative polyadenylation signal (TGTAA) for Chlamydomonas (Silflow et al, 1985) is located 207 base pairs downstream of the TAG stop codon. The UNI2 gene sequence is available at GenBank under the accession number EF675992.…”

Section: Cloning and Tagging The Uni2 Genementioning

confidence: 99%

The Uni2 Phosphoprotein is a Cell Cycle–regulated Component of the Basal Body Maturation Pathway inChlamydomonas reinhardtii

Piasecki

LaVoie

Tam

et al. 2008

MBoC

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Mutations in the UNI2 locus in Chlamydomonas reinhardtii result in a "uniflagellar" phenotype in which flagellar assembly occurs preferentially from the older basal body and ultrastructural defects reside in the transition zones. The UNI2 gene encodes a protein of 134 kDa that shares 20.5% homology with a human protein. Immunofluorescence microscopy localized the protein on both basal bodies and probasal bodies. The protein is present as at least two molecular-weight variants that can be converted to a single form with phosphatase treatment. Synthesis of Uni2 protein is induced during cell division cycles; accumulation of the phosphorylated form coincides with assembly of transition zones and flagella at the end of the division cycle. Using the Uni2 protein as a cell cycle marker of basal bodies, we observed migration of basal bodies before flagellar resorption in some cells, indicating that flagellar resorption is not required for mitotic progression. We observed the sequential assembly of new probasal bodies beginning at prophase. The uni2 mutants may be defective in the pathways leading to flagellar assembly and to basal body maturation. INTRODUCTIONCentrioles are evolutionarily ancient organelles, possibly extant in a common ancestor of all eukaryotic cells (CavalierSmith, 2002). They are required for the organization of interphase and mitotic microtubule arrays and as basal bodies they serve as templates for the growth of nine doublet microtubules in axonemes of cilia and flagella (for review see Marshall, 2007). Although flowering plants and more derived fungi have reduced or eliminated centrioles and cilia, metazoans have diversified their functions in numerous sensory and developmental processes (for reviews see Singla and Reiter, 2006;Dawe et al., 2007). Several mammalian genetic disorders have been linked to defects in centrioles or cilia (for review see Afzelius, 2004;Pan et al., 2005). For example, the proteins associated with Bardet-Biedl syndrome, a debilitating pleiotropic human disorder characterized by obesity, renal abnormalities, polydactyly, retinal degeneration, and genitourinary tract malformations have been shown to localize to or around centrioles and cilia (for review see Beales, 2005;Blacque and Leroux, 2006).Centrioles in actively dividing cells show conservative replication and semiconservative segregation during division (for review see Dawe et al., 2007). As a result, each pair of centrioles consists of an older and a younger centriole that differ in age by at least one cell generation. The "mother" centriole differs both in structure and function from the "daughter" centriole. For example, in the single pair of centrioles found in most types of mammalian cells, the mother centriole nucleates the growth of the primary cilium and contains proteins not present in the daughter centriole. The daughter centriole becomes competent for ciliary assembly only in the ensuing cell cycle (for review see Bornens, 2002). Extension of the pathway for sequential maturation of basal bodies has been ...

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“…Transcripts from all four genes appear to accumulate coordinately during flagellar regeneration (5,39,43), and sequence comparisons among these genes have revealed several conserved 5' flanking regions (4) as well as an intriguing homology among all the genes in one of their intervening sequences (38,43). These conserved sequences are potential control regions for the induction of tubulin mRNA synthesis during flagellar regeneration, but because tubulin is used by the cell in organelles other than the flagellum, such as the mitotic spindle, there is a possibility that these sequences function to regulate tubulin gene expression during cellular events other than the production of new flagella.…”

Section: Expression Of Radial Spoke and Dynein Genesmentioning

confidence: 99%

Molecular cloning and expression of flagellar radial spoke and dynein genes of Chlamydomonas.

Williams

Mitchell

Rosenbaum

1986

The Journal of Cell Biology

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Abstract. Several flagellar dynein ATPase and radial spokehead genes have been isolated from a Chlamydomonas genomic expression library in Lgtll. The library was probed with polyclonal and monoclonal antibodies raised against purified flagellar polypeptides, and recombinant phage giving positive signals were cloned. In vitro translation of mRNAs hybrid-selected by the cloned sequences from whole cell RNA provided confirmation of identity for three of the four clones. Evidence supporting the identification of the fourth, which encodes a dynein heavy chain, was provided by antibody selection; the fusion protein produced by this clone selected heavy chain-specific antibodies from a complex polyclonal antiserum recognizing many dynein determinants. One of the radial spoke sequences isolated here is of particular interest because it encodes the wild-type allele of a locus which was defined previously by temperature-sensitive paralyzed flagella mutation pf-26~ (Huang, B., G. Piperno, Z. Ramanis, and D. J. L. Luck, 1981, J. Cell Biol., 88:80-88). The cloned sequence was used to hybridselect mRNA from mutant pf-26~ cells, and when translated in vitro, the selected mRNA produced a mutant spokehead polypeptide with an altered electrophoretic mobility. This confirms that the pf-26~ mutation alters the primary structure of a radial spokehead polypeptide. To quantify spokehead and dynein mRNAs during flagellar regeneration, all of the cloned sequences were used as hybridization probes in RNA dot experiments. Levels increased rapidly and coordinately after deflagellation, peaked 3-10-fold above nondeflagellated controls, and then returned to control values within 2 h. This accumulation pattern was similar to that of flagellar ct-tubulin mRNA.

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The two alpha-tubulin genes of Chlamydomonas reinhardi code for slightly different proteins.

Cited by 174 publications

References 44 publications

Characterization of two genes encoding the mitochondrial alternative oxidase in Chlamydomonas reinhardtii

Characterization of two genes encoding the mitochondrial alternative oxidase in Chlamydomonas reinhardtii

The Uni2 Phosphoprotein is a Cell Cycle–regulated Component of the Basal Body Maturation Pathway inChlamydomonas reinhardtii

Molecular cloning and expression of flagellar radial spoke and dynein genes of Chlamydomonas.

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