Darkness and antibiotics increase the steady-state transcripts of the elongation factor gene (tuf) inChlamydomonas reinhardtii

Silk, Gregg W.; Wu, Madeline

doi:10.1007/bf00569335

Cited by 12 publications

(15 citation statements)

References 24 publications

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“…Exceptional increases were shown by clpP and tufA, particularly under nutrient stress. The tufA mRNA also exhibited increased abundance in the dark, as reported previously (Silk and Wu, 1988). Because clpP and tufA are involved in proteolysis and translation, respectively (Hwang et al, 1996;Majeran et al, 2000), these may be symptomatic of a post-transcriptional plastid stress response mechanism.…”

Section: R E T R a C T E Dsupporting

confidence: 81%

Retracted: The Chlamydomonas reinhardtii Organellar Genomes Respond Transcriptionally and Post-Transcriptionally to Abiotic Stimuli

2002

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The Chlamydomonas reinhardtii plastid and mitochondrial transcriptomes were surveyed for changes in RNA profiles resulting from growth in 12 culture conditions representing 8 abiotic stimuli. Organellar RNA abundance exhibited marked changes during nutrient stress and exposure to UV light, as revealed by both RNA gel blot and DNA microarray analyses. Of particular note were large increases in tufA and clpP transcript abundance during nutrient limitation. Phosphate and sulfur limitation resulted in the most global, yet opposite, effects on organellar RNA abundance, changes that were dissected further using run-on transcription assays. Removal of sulfate from the culture medium, which is known to reduce photosynthesis, resulted in 2-fold to 10-fold decreases in transcription rates, which were reflected in lower RNA abundance. The decrease in transcriptional activity was completely reversible and recovered to twice the control level after sulfate replenishment. Conversely, phosphate limitation resulted in a twofold to threefold increase in RNA abundance that was found to be a post-transcriptional effect, because it could be accounted for by increased RNA stability. This finding is consistent with the known metabolic slowdown under phosphate stress. Additionally, inhibitor studies suggested that unlike those in higher plants, Chlamydomonas chloroplasts lack a nucleusencoded plastid RNA polymerase. The apparently single type of polymerase could contribute to the rapid and genomewide transcriptional responses observed within the chloroplast.

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Section: R E T R a C T E Dsupporting

confidence: 81%

Retracted: The Chlamydomonas reinhardtii Organellar Genomes Respond Transcriptionally and Post-Transcriptionally to Abiotic Stimuli

2002

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“…The approximately 700-bp EcoRVHpal restriction fragment containing atpB protein-coding sequences (Woessner et al, 1986) was cloned into plasmid pUC7 to create plasmid pCrcatpB. The approximately 500-bp EcoRIPstl restriction fragment, which contains an internal portion of the tufA gene (Silk and Wu, 1988) was cloned into Bluescript vector to create plasmid pCrctufA. The approximately 520-bp Dral restriction fragment, which contains the rpll6 gene (Low et al, 1987) was cloned into pUC7 vector DNA to create plasmid pCrcrpll6.…”

Section: Plasmidsmentioning

confidence: 99%

Transcriptional Analysis of Endogenous and Foreign Genes in Chloroplast Transformants of Chlamydomonas

Blowers¹,

Ellmore²,

Klein³

et al. 1990

The Plant Cell

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Transcription from modified chloroplast genes has been studied in vitro, but only with the recently developed ability to stably introduce foreign DNA into Chlamydomonas reinhardtii chloroplast chromosomes in situ has it become possible to do so in vivo. Cloned chloroplast DNA sequences, into which had been inserted chimeric genes composed of the GUS coding sequence reporter under transcriptional control of chloroplast promoters for the C. reinhardtii atpA, atpB, and rbcL genes, were introduced into the cells on microprojectiles. These constructs become integrated into chloroplast chromosomes by homologous recombination. RNA gel blot analyses demonstrated that a single major 8-glucuronidase (GUS)-hybridizing transcript accumulates in each chloroplast transformant. We have found that: (1) Transcription of the chimeric gene begins at the same site as in the corresponding endogenous chloroplast gene; (2) the rates of transcription in vivo of the atpA:GUS and atpB:GUS genes relative to one another and to other genes are the same as those for the endogenous atpA and atpB genes, respectively, indicating that these promoters are fully functional despite being fused to a foreign gene and being at an alien location on the chloroplast chromosome; (3) in contrast to the atpA and atpB promoters, the rbcL promoter directs transcription of the rbcL:GUS gene at only 1% of the expected rate, suggesting that other features are required for optimal activity of this promoter; and (4) 22 base pairs upstream of the 5' end of the atpB:GUS transcript in the atpB promoter element is sufficient to confer wild-type levels of promoter activity.

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“…Cloning of the Cp psbA, rbcL, and 16S rRNA probes and the 3' end of tufA from Cp DNA plasmid libraries was previously reported [27]. P21, a 2.3 kb Pst I fragment containing the 5' end of the tufa gene and the 5'-flanking psbK gene [26] was cloned in Bluescript in both orientations to produce plasmids pP21 and pP21R respectively.…”

Section: Plasmids and General Cloning Proceduresmentioning

confidence: 99%

“…P21, a 2.3 kb Pst I fragment containing the 5' end of the tufa gene and the 5'-flanking psbK gene [26] was cloned in Bluescript in both orientations to produce plasmids pP21 and pP21R respectively. Deletions of these clones were constructed as previously described [27]. To construct plasmid pHDP47 (Fig.…”

Section: Plasmids and General Cloning Proceduresmentioning

confidence: 99%

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Posttranscriptional accumulation of chloroplast tufA (elongation factor gene) mRNA during chloroplast development in Chlamydomonas reinhardtii

Silk

1993

Plant Mol Biol

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Light induces chloroplast (Cp) differentiation in dark-grown y-1 strains of Chlamydomonas reinhardtii. Slot blot analysis was used to quantitate tufA, psbA, psbK, rbcL, and 16S rRNA transcript accumulation and transcription during Cp differentiation. When etiolated cc-125 y-1 cells were illuminated for 5 h, a 1710 bp tufA mRNA accumulated up to 5-fold while the psbA, rbcL, and 16S rRNA transcripts accumulated less than 1.5-fold. The tufA gene encodes translational elongation factor EF-Tu. The light-induced accumulation of tufA mRNA did not occur in cc-1931, a strain that does not become etiolated in darkness. Pulse labelling was used to measure the transcription of Cp transcripts during tufA mRNA accumulation, and no detectable change in tufA transcription was observed. These results imply that the half life of the tufA transcript increases during the greening process.

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Darkness and antibiotics increase the steady-state transcripts of the elongation factor gene (tuf) inChlamydomonas reinhardtii

Cited by 12 publications

References 24 publications

Retracted: The Chlamydomonas reinhardtii Organellar Genomes Respond Transcriptionally and Post-Transcriptionally to Abiotic Stimuli

Retracted: The Chlamydomonas reinhardtii Organellar Genomes Respond Transcriptionally and Post-Transcriptionally to Abiotic Stimuli

Transcriptional Analysis of Endogenous and Foreign Genes in Chloroplast Transformants of Chlamydomonas

Posttranscriptional accumulation of chloroplast tufA (elongation factor gene) mRNA during chloroplast development in Chlamydomonas reinhardtii

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