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Functional relationship of GUN1 and FUG1 in plastid proteostasis

ABSTRACT: We have previously identified the chloroplast translation initiation factor IF-2, or FUG1, as a GUN1 interactor based on co-immunoprecipitation and co-expression analysis (Tadini et al., 2016). To further investigate the functional relationship between these two proteins, the double mutant gun1-103 fug1-3 was generated by crossing the single gun1-103 knock-out mutant and the fug1-3 knock-down mutant. We performed comparative proteomics on gun1-103 and fug1-3, as well as on the double mutant gun1-103 fug1-3.

OTHER RELATED OMICS DATASETS IN: GSE123583

INSTRUMENT(S): impact II

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Seedling, Whole Body

SUBMITTER: Giada Marino

LAB HEAD: Dario Leister

PROVIDER: PXD012067 | Pride | 2019-06-26

REPOSITORIES: Pride

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Relationship of GUN1 to FUG1 in chloroplast protein homeostasis.

Marino Giada G Naranjo Belen B Wang Jing J Penzler Jan-Ferdinand JF Kleine Tatjana T Leister Dario D

The Plant journal : for cell and molecular biology 20190517 3

GUN1 integrates retrograde signals in chloroplasts but the underlying mechanism is elusive. FUG1, a chloroplast translation initiation factor, and GUN1 are co-expressed at the transcriptional level, and FUG1 co-immunoprecipitates with GUN1. We used mutants of GUN1 (gun1-103) and FUG1 (fug1-3) to analyse their functional relationship at the physiological and system-wide level, the latter including transcriptome and proteome analyses. Absence of GUN1 aggravates the effects of decreased FUG1 levels ...[more]

PMID: 31002470

Similar Datasets

Functional relationship of GUN1 and FUG1 in plastid proteostasis

Project description:GUN1 integrates retrograde signals in the chloroplast but the underlying mechanism is elusive. FUG1, a chloroplast translation initiation factor, and GUN1 are co-expressed at the transcript level, and FUG1 co-immunoprecipitates with GUN1. We used mutants of GUN1 (gun1-103) and FUG1 (fug1-3) to analyse their functional relationship at the physiological and systems-wide level, the latter including transcriptome and proteome analyses. Absence of GUN1 aggravates the effects of decreased FUG1 levels on chloroplast protein translation, resulting in transient additive phenotypes with respect to photosynthesis, leaf coloration, growth and cold acclimation. Variegation of the var2 mutant is enhanced by gun1-103 in terms of increasing the fraction of white sectors, in contrast to fug1-3 that acts as suppressor. The transcriptomes of fug1-3 and gun1-103 are very similar, but absence of GUN1 alone has almost no effects on protein levels, whereas chloroplast protein accumulation is markedly decreased in fug1-3. In gun1 fug1 double mutants, effects on transcriptomes and particularly proteomes are enhanced. Our results show that GUN1 function becomes critical when chloroplast proteostasis is perturbed by decreased translation (fug1) or degradation (var2) of chloroplast proteins. The functions of FUG1 and GUN1 appear to be related, corroborating the view that GUN1 operates in chloroplast proteostasis.

2019-04-23 | GSE123583 | GEO

pgr5 suppressor screen uncovers a distinct mechanism safeguarding the cytochrome b6f complex from damage through PGR5

Project description:PROTON GRADIENT5 (PGR5) is thought to promote cyclic electron flow (CEF) and its deficiency causes increased photosensitivity of photosystem I (PSI), leading to lethality under fluctuating light (FL). By screening for suppressor mutations that rescue FL lethality of pgr5 plants, we identified a portfolio of mutations affecting 12 photosynthesis-related proteins. Here we performed a proteomic analysis to compare a mutant lacking PGR5 vs wildtype ( at2g04360-1/pfsc1-1 vs. Col-0)

2024-03-15 | PXD047221 | Pride

ChIPseq of enterocytes of the jejunum from transgene C57BL/6 mice harboring a CKIαΔgut knock-out

Project description:ChIPseq of enterocytes of the jejunum from transgene C57BL/6 mice to assess the effects of a CKIαΔgut knock-out in the jejunum and a CKIαΔgut/p53 double knock-out. In addition, enterocytes expressing a mutant p53 (R172H) in combination with CKIαΔgut knockout were analyzed.

2020-02-25 | E-MTAB-7858 | biostudies-arrayexpress

Expression data from Arabidopsis wild-type, sicy-192, and sicy-192 gun1-101 plants

Project description:Plastid gene expression (PGE) acts as a signal that regulates the expression of photosynthesis-associated nuclear genes (PhANGs) via GENOMES UNCOUPLED1 (GUN1)-dependent retrograde signaling. We recently isolated Arabidopsis sugar-inducible cotyledon yellow-192 (sicy-192), a gain-of-function mutant of plastidic invertase (INV-E), and showed that following the treatment of this mutant with sucrose, the expression of PhANGs decreased while that of nitrate reductase 1 (NIA1) increased. Plastid-encoded RNA polymerase (PEP)-dependent PGE was markedly suppressed in the sicy-192 mutant by the sucrose treatment. A double mutant of sicy-192 and gun1-101, a null mutant of GUN1, revealed that metabolic perturbation in the sucrose-treated sicy-192 mutant was, at least in part, dependent on GUN1. To confirm whether there is a relationship between plastid sugar metabolism and nuclear gene expression, we performed a microarray analysis using Suc-treated 3 genotypes consisting of wild-type, sicy-192, and sicy-192 gun1-101 plants. Wild-type, sicy-192, and sicy-192 gun1-101 plants were grown on MS medium containing 100 mM sucrose in a growth chamber kept at 25Â°C during 16 h of light (100 Âµmol/m2/s) and at 22Â°C during 8 h of darkness for 4 days. At 4 days of age, cotyledons were collected as samples for microarray analysis.

2013-09-14 | E-GEOD-50857 | biostudies-arrayexpress

Root transcriptome of the tsn1 tsn2 double mutant

Project description:au06-03_tsn_fj - tsn_fj - Root transcriptome of the tsn1 tsn2 double mutant - Comparison of a tsn1 tsn2 double mutant line (A10.9.3- or A13.3.11-) and an isogenic line complemented by TSN2 (A10.9.5) or TSN1 (A13.3.2) Keywords: gene knock in (transgenic),gene knock out

2009-04-08 | GSE15559 | GEO

Genome-wide hsa-miR-503, hsa-miR-103, and hsa-miR-494 target profiles [array]

Project description:Gene expression profile following transfection with miR-503, miR-103, or miR-494 mature duplex Examination of mRNA levels in HeLa cells following transfection of miR-503, miR-103, or miR-494 mature duplex, control siRNA against GFP, or mock transfection (lipofectamine 2000 alone)

2015-02-04 | E-GEOD-64364 | biostudies-arrayexpress

Expression data from Arabidopsis wild-type, sicy-192, and sicy-192 gun1-101 plants

2013-09-14 | GSE50857 | GEO

Nontargeted and targeted metabolomics measurements of abiotic stress responses in three-week-old Arabidopsis thaliana plants' rosette leaf tissue

Project description:Nontargeted and targeted metabolomics measurements of abiotic stress responses in three-week-old Arabidopsis thaliana plants' rosette leaf tissue for Col-0 wild type plants and double/triple knockout mutants of aquaporins (pip2;1 pip2;2 and pip2;1 pip2;2 pip2;4) treated with drought, heat at different air humidities, or combined drought-heat stress at different air humidities. This experiment contains FT-ICR-MS measurements for 103 Arabidopsis thaliana rosette leaf samples covering three genotypes under six different environmental conditions. The three genotypes comprise the Col-0 wildtype and two loss-of-function mutants of aquaporins, a pip2;1 pip2;2 double mutant and a pip2;1 pip2;2 pip2;4 triple mutant (respective AGI locus identifiers: AT3G53420, AT2G37170, AT5G60660). The six conditions include control condition (well-watered, 22 °C, 70% relative air humidity), drought stress (one week without watering), heat stress without changing the absolute humidity of the ambient air (6 hours at 33 °C, 37% relative air humidity), heat stress with supplemented air humidity to maintain a constant vapor pressure deficit before and during the heat episode (6 hours at 33 °C, 84% relative air humidity), and the combinations of drought pretreatment with each of the two heat stress variants (one week of drought followed by 6 hours of heat stress). Samples from all conditions were harvested at the same time (within 15 min starting at 5 pm). For validation, GC-TOF-MS measurements were done for two genotypes (wildtype, double mutant) and two conditions (drought, control) on partially overlapping samples.

2017-07-17 | MTBLS355 | MetaboLights

ntr-Transcriptomic study of a thioredoxin reductase knock-out mutant.

Project description:rs04-02_ntr - ntr - Thioredoxins are small redox proteins implicated in crucial pathways of cell life. They are reduced by a conserved flavoprotein named NADPH-dependent thioredoxin reductase (NTR). In order to give an insight into the functions of the NTR/thioredoxin pathway in Arabidopsis, we have used a reverse genetic approch. Both NTR genes (NTRA and NTRB) have been inactivated in the double ntra ntrb mutant. T-DNA lines have been provided by the SALK library. Surprisingly, this mutant shows a limited phenotype suggested that some additional redox pathways are involved in compensating the inactivation of the NTR/thioredoxin pathway. In order to isolate compensatory genes that would be transcriptionally induced in the ntra ntrb mutant, we have performed this CATMA project. - Comparison of the transcript profile of the ntra ntrb knock-out mutant vs Col-0 wild-type ecotype. Seeds of the wild-type Col-0 ecotype and of the ntra ntrb homozygote double mutant were sowed in soil. Three replicates of lines were made (Col-0 1, 2 and 3 ntra ntrb 1, 2 and 3) : plants were put at different places of the greenhouse and grown at different times. Germination was synchronized by 4 days at 4degreeC and plants were grown during 17 days, until they reached a 6 rosette leaves stage. Entire plants (30 plants per sample) were harvested, frozen in liquid nitrogen and stored at 80degreeC until extraction. Keywords: gene knock out,wt vs mutant comparison 3 dye-swap - CATMA arrays

2007-06-19 | E-GEOD-7998 | biostudies-arrayexpress

Rhizobium leguminosarum glutathione peroxidase is essential for oxidative stress resistance and efficient nodulation

Project description:Glutathione plays a tremendous role in regulating the homeostasis of redox state, and appears to be essential for proper development of the root nodules. Glutathione peroxidase catalyzes the reduction of peroxides by oxidation of GSH to oxidized GSH (GSSG), which can in turn be reduced by glutathione reductase (GR). However, it has not been determined whether the R. leguminosarum Gpx or GR is required during symbiotic interactions with pea. To characterise the role of glutathione-dependent enzymes in the symbiotic process, single and double mutants were made in gpxA and gorA genes. All the mutations did not affect the growth of R. leguminosarum, but they increased the sensitivity of R. leguminosarum strains to H2O2. The gorA mutant can induce the formation of normal nodules, while the gpxA single and double mutants exhibited an abnormal nodulation phenotype coupled to more than 50% reduction in nitrogen-fifixing capacity, this defect in nodulation was characterized by the formation of undeveloped and ineffective nodules. In addition, the gorA and gpxA double mutant was severely impaired in rhizosphere colonization. LC-MS/MS analysis quantitative proteomics techniques were employed to compare differential gpxA mutant root bacteroids in response to the wild type infection. Sixty differentially expressed proteins were identified including seven up-regulated and twenty down-regulated proteins. By sorting the down-regulated proteins according to metabolic function, eight proteins were transporter protein, seven proteins were dehydrogenases and deoxygenases. Moreover, three down-regulation proteins are directly involved in nodule process.

2020-11-16 | PXD022029 | Pride

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