scholarly journals Multiple Translational Control Sequences in the 5′ Leader of the Chloroplast psbC mRNA Interact With Nuclear Gene Products in Chlamydomonas reinhardtii

Genetics ◽  
2003 ◽  
Vol 163 (3) ◽  
pp. 895-904
Author(s):  
William Zerges ◽  
Andrea H Auchincloss ◽  
Jean-David Rochaix
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Reporter Gene ◽  
Translational Control ◽  
Nuclear Gene ◽  
Gene Products ◽  
Wild Type ◽  
Translation Initiation Region ◽  
Mrna Stabilization ◽  
In The Wild ◽  
Nuclear Loci

Abstract Translation of the chloroplast psbC mRNA in the unicellular eukaryotic alga Chlamydomonas reinhardtii is controlled by interactions between its 547-base 5′ untranslated region and the products of the nuclear loci TBC1, TBC2, and possibly TBC3. In this study, a series of site-directed mutations in this region was generated and the ability of these constructs to drive expression of a reporter gene was assayed in chloroplast transformants that are wild type or mutant at these nuclear loci. Two regions located in the middle of the 5′ leader and near the initiation codon are important for translation. Other deletions still allow for partial expression of the reporter gene in the wild-type background. Regions with target sites for TBC1 and TBC2 were identified by estimating the residual translation activity in the respective mutant backgrounds. TBC1 targets include mostly the central part of the leader and the translation initiation region whereas the only detected TBC2 targets are in the 3′ part. The 5′-most 93 nt of the leader are required for wild-type levels of transcription and/or mRNA stabilization. The results indicate that TBC1 and TBC2 function independently and further support the possibility that TBC1 acts together with TBC3.

scholarly journals Translation of the chloroplast psbC mRNA is controlled by interactions between its 5' leader and the nuclear loci TBC1 and TBC3 in Chlamydomonas reinhardtii.

1997 ◽  
Vol 17 (6) ◽  
pp. 3440-3448 ◽  
Author(s):  
W Zerges ◽  
J Girard-Bascou ◽  
J D Rochaix
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Translation Initiation ◽  
Translational Control ◽  
Reporter Genes ◽  
Activation Function ◽  
Wild Type ◽  
Cis Acting ◽  
Translation Initiation Codon ◽  
Nuclear Loci

Translation of the chloroplast psbC mRNA in Chlamydomonas reinhardtii has been shown previously to require interactions between its 5' untranslated region (5' UTR) and the functions encoded by two nuclear loci, which we name here TBC1 and TBC2. We show that a 97-nucleotide (nt) region located in the middle of the psbC 5' UTR is required for translation initiation. Unlike most procaryotic cis-acting translational control elements, this region has a translational activation function and is located 236 nt upstream from the GUG translation initiation codon. In vivo pulse-labeling of chloroplast-encoded proteins and analyses of the expression of chimeric reporter genes in vivo reveal that a mutation of a newly described locus, TBC3, restores translation from the psbC 5' UTR in the absence of either this cis-acting element or the wild-type trans-acting TBC1 function. These data demonstrate that sequences located in the middle of the psbC 5' UTR, TBC1, and TBC3 functionally interact to control the translation of the psbC mRNA.

scholarly journals Atp10p Assists Assembly of Atp6p into the F0Unit of the Yeast Mitochondrial ATPase

2004 ◽  
Vol 279 (19) ◽  
pp. 19775-19780 ◽  
Author(s):  
Alexander Tzagoloff ◽  
Antoni Barrientos ◽  
Walter Neupert ◽  
Johannes M. Herrmann
Keyword(s):  
Nuclear Gene ◽  
Mitochondrial Atpase ◽  
Gene Products ◽  
Wild Type ◽  
Mitochondrial Translation ◽  
Mitochondrial Ribosomes ◽  
Mitochondrial Inner Membrane ◽  
Atpase Subunits ◽  
Specific Factors

The F0F1-ATPase complex of yeast mitochondria contains three mitochondrial and at least 17 nuclear gene products. The coordinate assembly of mitochondrial and cytosolic translation products relies on chaperones and specific factors that stabilize the pools of some unassembled subunits. Atp10p was identified as a mitochondrial inner membrane component necessary for the biogenesis of the hydrophobic F0sector of the ATPase. Here we show that, following its synthesis on mitochondrial ribosomes, subunit 6 of the ATPase (Atp6p) can be cross-linked to Atp10p. This interaction is required for the integration of Atp6p into a partially assembled subcomplex of the ATPase. Pulse labeling and chase of mitochondrial translation productsin vivoindicate that Atp6p is less stable and more rapidly degraded in anatp10null mutant than in wild type. Based on these observations, we propose Atp10p to be an Atp6p-specific chaperone that facilitates the incorporation of Atp6p into an intermediate subcomplex of ATPase subunits.

scholarly journals Physiological response of two different Chlamydomonas reinhardtii strains to light–dark rhythms

Botany ◽  
2016 ◽  
Vol 94 (1) ◽  
pp. 53-64 ◽  
Author(s):  
Michael Sandmann ◽  
Andreas Garz ◽  
Ralf Menzel
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Cell Volume ◽  
Dark Respiration ◽  
Starch Content ◽  
Saturation Index ◽  
Cell Physiology ◽  
Wild Type ◽  
Cell Productivity ◽  
In The Wild ◽  
Entire Cell

Cells of a cell-wall deficient line (cw15-type) of Chlamydomonas reinhardtii and of the corresponding wild type were grown during repetitive light–dark cycles. In a direct comparison, both lines showed approximately the same relative biomass increase during light phase but the cw-line produced significantly more, and smaller, daughter cells. Throughout the light period the average cellular starch content, the cellular chlorophyll content, the cellular rate of dark respiration, and the cellular rate of photosynthesis of the cw-line was lower. Despite this, several non-cell volume related parameters like the development of starch content per cell volume were clearly different over time between the strains. Additionally, the chlorophyll-based photosynthesis rates were 2-fold higher in the mutant than in the wild-type cells, and the ratio of chlorophyll a to chlorophyll b as well as the light-saturation index were also consistently higher in the mutant cells. Differences in the starch content were also confirmed by single cell analyses using a sensitive SHG-based microscopy approach. In summary, the cw15-type mutant deviates from its genetic background in the entire cell physiology. Both lines should be used in further studies in comparative systems biology with focus on the detailed relation between cell volume increase, photosynthesis, starch metabolism, and daughter cell productivity.

scholarly journals Suppression of a defect in the 5' untranslated leader of mitochondrial COX3 mRNA by a mutation affecting an mRNA-specific translational activator protein.

1993 ◽  
Vol 13 (8) ◽  
pp. 4806-4813 ◽  
Author(s):  
M C Costanzo ◽  
T D Fox
Keyword(s):  
Nuclear Gene ◽  
Gene Products ◽  
Wild Type ◽  
Large Deletions ◽  
Site Of Action ◽  
Genes Encoding ◽  
Cold Sensitive ◽  
Translational Activator ◽  
A Site ◽  
Dominant Suppressor

Translation of the Saccharomyces cerevisiae mitochondrial COX3 mRNA, encoding subunit III of cytochrome c oxidase, specifically requires the action of the nuclear gene products PET54, PET122, and PET494 at a site encoded in the 612-base 5' untranslated leader. To identify more precisely the site of action of the translational activators, we constructed two large deletions of the COX3 mRNA 5' untranslated leader. Both deletions blocked translation without affecting mRNA stability. However, one of the large deletions was able to revert to partial function by a small secondary deletion within the remaining 5' leader sequences. Translation of the resulting mutant (cox3-15) mRNA was still dependent on the nuclear-encoded specific activators but was cold sensitive. We selected revertants of this mitochondrial mutant at low temperature to identify genes encoding proteins that might interact with the COX3 mRNA 5' leader. One such revertant carried a missense mutation in the PET122 gene that was a strong and dominant suppressor of the cold-sensitive defect in the mRNA, indicating that the PET122 protein interacts functionally (possibly directly) with the COX3 mRNA 5' leader. The cox3-15 mutation was not suppressed by overproduction of the wild-type PET122 protein but was very weakly suppressed by overproduction of PET494 and slightly better suppressed by co-overproduction of PET494 and PET122.

Characterization of a norflurazon-resistant mutant of Chlamydomonas reinhardtii

Weed Science ◽  
1997 ◽  
Vol 45 (3) ◽  
pp. 374-377 ◽  
Author(s):  
Varsha Vartak ◽  
Sujata Bhargava
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Mode Of Action ◽  
Resistant Mutant ◽  
Phytoene Desaturase ◽  
Wild Type ◽  
Cross Tolerance ◽  
In The Wild ◽  
Similar Mode ◽  
Target Enzyme

A norflurazon-resistant mutant has been isolated from Chlamydomonas reinhardtii that showed a three-fold factor of resistance over wild type cultures. In comparison to wild type cultures, the mutant showed better retention of chlorophylls and carotenoids when grown in light in the presence of norflurazon. When grown in the dark, chlorophyll losses were similar, while carotenoid losses were lower than in the wild type cultures. Higher levels of phytoene accumulated in the wild type cultures in the presence of norflurazon than in the resistant cultures. The resistant cultures also showed cross tolerance to EMD-IT 5914, a herbicide with a similar mode of action. Norflurazon resistance in this alga appears to arise from alterations in the target enzyme phytoene desaturase.

scholarly journals Exopolyphosphatases PPX1 and PPX2 from Corynebacterium glutamicum

2009 ◽  
Vol 75 (10) ◽  
pp. 3161-3170 ◽  
Author(s):  
Steffen N. Lindner ◽  
Sandra Knebel ◽  
Hendrik Wesseling ◽  
Siegfried M. Schoberth ◽  
Volker F. Wendisch
Keyword(s):  
Corynebacterium Glutamicum ◽  
Type Strain ◽  
Wild Type Strain ◽  
Deletion Mutants ◽  
Gene Products ◽  
Wild Type ◽  
Inorganic Polyphosphate ◽  
Deletion Mutations ◽  
In The Wild ◽  
Growth Experiments

ABSTRACT Corynebacterium glutamicum accumulates up to 300 mM of inorganic polyphosphate (PolyP) in the cytosol or in granules. The gene products of cg0488 (ppx1) and cg1115 (ppx2) were shown to be active as exopolyphosphatases (PPX), as overexpression of either gene resulted in higher exopolyphosphatase activities in crude extracts and deletion of either gene with lower activities than those of the wild-type strain. PPX1 and PPX2 from C. glutamicum share only 25% identical amino acids and belong to different protein groups, which are distinct from enterobacterial, archaeal, and yeast exopolyphosphatases. In comparison to that in the wild type, more intracellular PolyP accumulated in the Δppx1 and Δppx2 deletion mutations but less when either ppx1 or ppx2 was overexpressed. When C. glutamicum was shifted from phosphate-rich to phosphate-limiting conditions, a growth advantage of the deletion mutants and a growth disadvantage of the overexpression strains compared to the wild type were observed. Growth experiments, exopolyphosphatase activities, and intracellular PolyP concentrations revealed PPX2 as being a major exopolyphosphatase from C. glutamicum. PPX2His was purified to homogeneity and shown to be active as a monomer. The enzyme required Mg2+ or Mn2+ cations but was inhibited by millimolar concentrations of Mg2+, Mn2+, and Ca2+. PPX2 from C. glutamicum was active with short-chain polyphosphates, even accepting pyrophosphate, and was inhibited by nucleoside triphosphates.

scholarly journals TheS. cerevisiaeHAP Complex, a Key Regulator of Mitochondrial Function, Coordinates Nuclear and Mitochondrial Gene Expression

2003 ◽  
Vol 4 (1) ◽  
pp. 37-46 ◽  
Author(s):  
S. Buschlen ◽  
J-M Amillet ◽  
B. Guiard ◽  
A. Fournier ◽  
C. Marcireau ◽  
...  
Keyword(s):  
Gene Expression ◽  
Respiratory Chain ◽  
Mitochondrial Gene ◽  
Nuclear Gene ◽  
Negative Control ◽  
Nuclear Genes ◽  
Wild Type ◽  
Mitochondrial Translation ◽  
Respiratory Chain Complexes ◽  
In The Wild

We have comparedSaccharomyces cerevisiaeglobal gene expression in wild-type and mutants (Δhap2 and Δhap4) of the HAP transcriptional complex, which has been shown to be necessary for growth on respiratory substrates. Several hundred ORFs are under positive or negative control of this complex and we analyse here in detail the effect of HAP on mitochondria. We found that most of the genes upregulated in the wild-type strain were involved in organelle functions, but practically none of the downregulated ones. Nuclear genes encoding the different subunits of the respiratory chain complexes figure in the genes more expressed in the wild-type than in the mutants, as expected, but in this group we also found key components of the mitochondrial translation apparatus. This control of mitochondrial translation may be one of the means of coordinating mitochondrial and nuclear gene expression in elaborating the respiratory chain. In addition, HAP controls the nuclear genes involved in several other mitochondrial processes (import, mitochondrial division) that define the metabolic state of the cell, but not mitochondrial DNA replication and transcription. In most cases, a putative CCAAT-binding site is present upstream of the ORF, while in others no such sites are present, suggesting the control to be indirect. The large number of genes regulated by the HAP complex, as well as the fact that HAP also regulates some putative transcriptional activators of unknown function, place this complex at a hierarchically high position in the global transcriptional regulation of the cell.

scholarly journals TCA1, a Single Nuclear-Encoded Translational Activator Specific for petA mRNA in Chlamydomonas reinhardtii Chloroplast

Genetics ◽  
2001 ◽  
Vol 159 (1) ◽  
pp. 119-132
Author(s):  
K Wostrikoff ◽  
Y Choquet ◽  
F-A Wollman ◽  
J Girard-Bascou
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Nuclear Gene ◽  
Photosynthetic Electron Transport ◽  
Cytochrome F ◽  
Coding Region ◽  
Reading Frame ◽  
Transport Chain ◽  
Nuclear Loci ◽  
Translational Activator ◽  
B6f Complex

Abstract We isolated seven allelic nuclear mutants of Chlamydomonas reinhardtii specifically blocked in the translation of cytochrome f, a major chloroplast-encoded subunit of the photosynthetic electron transport chain encoded by the petA gene. We recovered one chloroplast suppressor in which the coding region of petA was now expressed under the control of a duplicated 5′ untranslated region from another open reading frame of presently unknown function. Since we also recovered 14 nuclear intragenic suppressors, we ended up with 21 alleles of a single nuclear gene we called TCA1 for translation of cytochrome b6f complex petA mRNA. The high number of TCA1 alleles, together with the absence of genetic evidence for other nuclear loci controlling translation of the chloroplast petA gene, strongly suggests that TCA1 is the only trans-acting factor. We studied the assembly-dependent regulation of cytochrome f translation—known as the CES process—in TCA1-mutated contexts. In the presence of a leaky tca1 allele, we observed that the regulation of cytochrome f translation was now exerted within the limits of the restricted translational activation conferred by the altered version of TCA1 as predicted if TCA1 was the ternary effector involved in the CES process.

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