Dataset Information

A systems biology-oriented investigation of Arabidopsis retrograde-signaling-defective mutant gun1 reveals the co-expression-based HUBs at the center of plastid development and homeostasis

ABSTRACT: In the present study we investigated at the system level the proteome obtained from seedlings of Arabidopsis thaliana wild-type Col-0 and gun1-101 and gun1-102 mutant plants treated or not with lincomycin (Lin), a chloroplast translation inhibitor widely used to inhibit chloroplast biogenesis and to trigger the plastid-to-nucleus retrograde communication. Specifically, we attempted a description of cellular responses to Lincomycin in wild-type genetic background and in retrograde signaling-defective mutant gun1. Hypotheses and key players emerged by our holistic view were validated at the experimental level. Our findings suggest that, in the presence of Lincomycin, cell responses rely on the plastid compartment in a GUN1-dependent manner, while in the gun1-mutant background, the activity of extra-plastid compartments prevails. Moreover, results obtained by complementary approaches depict the Oxygen Evolving Complex subunit PsbO as an atypical Photosynthesis-related protein by accumulating in non-photosynthetic plastids and having a superhub role in modulating chloroplast dismantling upon impairment of plastid functions.

INSTRUMENT(S):

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

TISSUE(S): Plant Cell, Shoot

SUBMITTER: Milena Marsoni

LAB HEAD: Guido Domingo

PROVIDER: PXD051970 | Pride | 2025-07-14

REPOSITORIES: Pride

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Publications

A Holistic Investigation of Arabidopsis Proteomes Altered in Chloroplast Biogenesis and Retrograde Signalling Identifies PsbO as a Key Regulator of Chloroplast Quality Control.

Di Silvestre Dario D Jeran Nicolaj N Domingo Guido G Vannini Candida C Marsoni Milena M Fortunato Stefania S de Pinto Maria Concetta MC Tamborrino Alberto A Negroni Yuri Luca YL Zottini Michela M Hong Lien Tran LT Lomagno Andrea A Mauri Pierluigi P Pesaresi Paolo P Tadini Luca L

Plant, cell & environment 20250514 8

Communication between the diverse compartments of plant cells relies on an intricate network of molecular interactions that orchestrate organellar development and adaptation to environmental conditions. Plastid-to-nucleus signalling pathways play a key role in relaying information from developing, mature, and damaged or disintegrating chloroplasts to the nucleus, which serves to coordinate gene expression between the two genomes. To shed light on these mechanisms, we performed a comprehensive an ...[more]

PMID: 40366233

Similar Datasets

Project description:Regulation of expression of genes encoding chloroplast components is critical to the autotrophic plant and never more so than in the cotyledons of the de-etiolating seedling. Many chloroplast proteins are nuclear-encoded and a retrograde signal from the chloroplasts (the Plastid Signal) modulates nuclear transcription. However, not all chloroplast-targeted genes are subject to this control and not all plastid-dependent nuclear genes are chloroplast-targeted. We therefore aim to provide the most comprehensive screen yet of which genes are affected by plastid-signalling. To specifically knock-out positive plastid signalling in light-grown cotyledons, the herbicide Norflurazon (NF) is supplied in the growth medium, causing a carotenoid deficiency that leaves the chloroplasts vulnerable to photobleaching. This blocks the expression of a subset of nuclear genes, such as Lhcb and HEMA1. Two pairs of RNAs will directly compare the transcription in seedlings grown under continuous white light with and without NF. A third RNA will also be compared from a mutant that shows a degree of constitutive positive plastid signalling. These two mutants act synergistically to counteract the effect of NF on nuclear transcription. The gun1,gun5 double mutant maintains a significantly higher level of Lhcb and HEMA1 expression in the presence of photobeached chloroplasts than the NF-treated wild-type. This transcriptome set will therefore complement RNA1 (wild-type+NF) and indicate which of the genes identified from the RNA1/RNA2 comparison are subject to the particular gun1/gun5 plastid signalling pathway(s). The growth of conditions of the seedlings (namely on MS medium supplemented with 1.5% sucrose, for 3 days under continuous WL following 2 days germination in darkness) has been chosen from the results of our own recent studies using Northern blotting techniques that show these conditions to maximise the respective NF and gun mutant effects on Lhcb and HEMA1 gene expression. This experiment is part one of a two-part study to compare the transcriptional output of this NF-affected pathway with that of a newly discovered FR-mediated pathway. A subsequent array experiment will assess the nuclear response as affected by this FR/ phyA-input pathway and the two sets of array data will be compared and contrasted. Note: Col-0 wild-type (NASC code N1092). gun1,gun5 double mutant(obtained from Enriquez Lopez-Juez, Royal Holloway, University of London.(Mochizuki et al. 2001 PNAS 98: 2053-2058). This line cannot be donated by us as it is the IP of Joanne Chory (SALK Institute, USA). Treatment = a herbicide (Norflurazon) application which leads to chloroplast photobleaching and hence down regulation of nuclear genes dependent on plastid signalling from intact chloroplasts.

Project description:Regulation of expression of genes encoding chloroplast components is critical to the autotrophic plant and never more so than in the cotyledons of the de-etiolating seedling. Many chloroplast proteins are nuclear-encoded and a retrograde signal from the chloroplasts (the Plastid Signal) modulates nuclear transcription. However, not all chloroplast-targeted genes are subject to this control and not all plastid-dependent nuclear genes are chloroplast-targeted. We therefore aim to provide the most comprehensive screen yet of which genes are affected by plastid-signalling. To specifically knock-out positive plastid signalling in light-grown cotyledons, the herbicide Norflurazon (NF) is supplied in the growth medium, causing a carotenoid deficiency that leaves the chloroplasts vulnerable to photobleaching. This blocks the expression of a subset of nuclear genes, such as Lhcb and HEMA1. Two pairs of RNAs will directly compare the transcription in seedlings grown under continuous white light with and without NF. A third RNA will also be compared from a mutant that shows a degree of constitutive positive plastid signalling. These two mutants act synergistically to counteract the effect of NF on nuclear transcription. The gun1,gun5 double mutant maintains a significantly higher level of Lhcb and HEMA1 expression in the presence of photobeached chloroplasts than the NF-treated wild-type. This transcriptome set will therefore complement RNA1 (wild-type+NF) and indicate which of the genes identified from the RNA1/RNA2 comparison are subject to the particular gun1/gun5 plastid signalling pathway(s). The growth of conditions of the seedlings (namely on MS medium supplemented with 1.5% sucrose, for 3 days under continuous WL following 2 days germination in darkness) has been chosen from the results of our own recent studies using Northern blotting techniques that show these conditions to maximise the respective NF and gun mutant effects on Lhcb and HEMA1 gene expression. This experiment is part one of a two-part study to compare the transcriptional output of this NF-affected pathway with that of a newly discovered FR-mediated pathway. A subsequent array experiment will assess the nuclear response as affected by this FR/ phyA-input pathway and the two sets of array data will be compared and contrasted. Note:; Col-0 wild-type (NASC code N1092). gun1,gun5 double mutant(obtained from Enriquez Lopez-Juez, Royal Holloway, University of London.(Mochizuki et al. 2001 PNAS 98: 2053-2058). This line cannot be donated by us as it is the IP of Joanne Chory (SALK Institute, USA). Treatment = a herbicide (Norflurazon) application which leads to chloroplast photobleaching and hence down regulation of nuclear genes dependent on plastid signalling from intact chloroplasts. Experimenter name = Alex McCormac; Experimenter phone = 023 80594297; Experimenter fax = 023 80594319; Experimenter address = School of Biological Sciences; Experimenter address = Division of Cell Sciences; Experimenter address = University of Southampton; Experimenter address = Bassett Crescent East; Experimenter address = Southampton; Experimenter zip/postal_code = SO16 7PX; Experimenter country = UK Experiment Overall Design: 6 samples were used in this experiment

			Action	DRS
	20161010_02_Gun_con3.raw	Raw
	20161010_03_WT_Lyn3.raw	Raw
	20161010_04_WT_Lyn1.raw	Raw
	20161010_05_Gun_Lyn3.raw	Raw
	20161010_07_Gun_Lyn2.raw	Raw