Proteomic analysis of haem-binding protein from
            Arabidopsis thaliana
            and
            Cyanidioschyzon merolae

Chloroplast biogenesis involves the coordinated expression of the plastid and nuclear genomes, requiring information to be sent from the nucleus to the developing chloroplasts and vice versa. Although it is well known how the nucleus controls chloroplast development, it is still poorly understood how the plastid communicates with the nucleus. Currently, haem is proposed as a plastid-to-nucleus (retrograde) signal that is involved in various physiological regulations, such as photosynthesis-associated nuclear genes expression and cell cycle in plants and algae. However, components that transduce haem-dependent signalling are still unidentified. In this study, by using haem-immobilized high-performance affinity beads, we performed proteomic analysis of haem-binding proteins from Arabidopsis thaliana and Cyanidioschyzon merolae . Most of the identified proteins were non-canonical haemoproteins localized in various organelles. Interestingly, half of the identified proteins were nucleus proteins, some of them have a similar function or localization in either or both organisms. Following biochemical analysis of selective proteins demonstrated haem binding. This study firstly demonstrates that nucleus proteins in plant and algae show haem-binding properties. This article is part of the theme issue ‘Retrograde signalling from endosymbiotic organelles’.

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AtRsgA from Arabidopsis thaliana controls maturation of the small subunit of the chloroplast ribosome

10.1101/225052 ◽

2017 ◽

Cited By ~ 1

Author(s):

Marcin Janowski ◽

Reimo Zoschke ◽

Lars Scharff ◽

Silvia Martinez Jaime ◽

Camilla Ferrari ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Proteomic Analysis ◽

Ribosome Biogenesis ◽

Functional Characterization ◽

Small Subunit ◽

Signalling Pathways ◽

Compensatory Mechanism ◽

Transcriptional Responses ◽

Retrograde Signalling ◽

Assembly Factor

SummaryPlastid ribosomes are very similar in structure and function to ribosomes of their bacterial ancestors. Since ribosome biogenesis is not thermodynamically favourable at biological conditions, it requires activity of many assembly factors. Here, we have characterized a homolog of bacterial rsgA in Arabidopsis thaliana and show that it can complement the bacterial homolog. Functional characterization of a strong mutant in Arabidopsis revealed that the protein is essential for plant viability, while a weak mutant produced dwarf, chlorotic plants that incorporated immature pre-16S ribosomal RNA into translating ribosomes. Physiological analysis of the mutant plants revealed smaller, but more numerous chloroplasts in the mesophyll cells, reduction of chlorophyll a and b, depletion of proplastids from the rib meristem and decreased photosynthetic electron transport rate and efficiency. Comparative RNA-sequencing and proteomic analysis of the weak mutant and wild-type plants revealed that various biotic stress-related, transcriptional regulation and post-transcriptional modification pathways were repressed in the mutant. Intriguingly, while nuclear- and chloroplast-encoded photosynthesis-related proteins were less abundant in the mutant, the corresponding transcripts were upregulated, suggesting an elaborate compensatory mechanism, potentially via differentially active retrograde signalling pathways. To conclude, this study reveals a new chloroplast ribosome assembly factor and outlines the transcriptomic and proteomic responses of the compensatory mechanism activated during decreased chloroplast function.Significance statementAtRsgA is an assembly factor necessary for maturation of the small subunit of the chloroplast ribosome. Depletion of AtRsgA leads to dwarfed, chlorotic plants and smaller, but more numerous chloroplasts. Large-scale transcriptomic and proteomic analysis revealed that chloroplast-encoded and - targeted proteins were less abundant, while the corresponding transcripts were upregulated in the mutant. We analyse the transcriptional responses of several retrograde signalling pathways to suggest a mechanism underlying this compensatory response.

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Identification and Quantification of Coumarins by UHPLC-MS in Arabidopsis Thaliana Natural Populations

Molecules ◽

10.3390/molecules26061804 ◽

2021 ◽

Vol 26 (6) ◽

pp. 1804

Author(s):

Izabela Perkowska ◽

Joanna Siwinska ◽

Alexandre Olry ◽

Jérémy Grosjean ◽

Alain Hehn ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

High Performance ◽

Biological Activities ◽

Natural Populations ◽

Biologically Active ◽

Stress Factors ◽

Mass Spectrometry Analysis ◽

Engineering Research ◽

Spectrometry Analysis ◽

Model Plant Arabidopsis Thaliana

Coumarins are phytochemicals occurring in the plant kingdom, which biosynthesis is induced under various stress factors. They belong to the wide class of specialized metabolites well known for their beneficial properties. Due to their high and wide biological activities, coumarins are important not only for the survival of plants in changing environmental conditions, but are of great importance in the pharmaceutical industry and are an active source for drug development. The identification of coumarins from natural sources has been reported for different plant species including a model plant Arabidopsis thaliana. In our previous work, we demonstrated a presence of naturally occurring intraspecies variation in the concentrations of scopoletin and its glycoside, scopolin, the major coumarins accumulating in Arabidopsis roots. Here, we expanded this work by examining a larger group of 28 Arabidopsis natural populations (called accessions) and by extracting and analysing coumarins from two different types of tissues–roots and leaves. In the current work, by quantifying the coumarin content in plant extracts with ultra-high-performance liquid chromatography coupled with a mass spectrometry analysis (UHPLC-MS), we detected a significant natural variation in the content of simple coumarins like scopoletin, umbelliferone and esculetin together with their glycosides: scopolin, skimmin and esculin, respectively. Increasing our knowledge of coumarin accumulation in Arabidopsis natural populations, might be beneficial for the future discovery of physiological mechanisms of action of various alleles involved in their biosynthesis. A better understanding of biosynthetic pathways of biologically active compounds is the prerequisite step in undertaking a metabolic engineering research.

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Protein Preparation for Proteomic Analysis of the Unfolded Protein Response in Arabidopsis thaliana

Methods in Molecular Biology - The Unfolded Protein Response ◽

10.1007/978-1-0716-1732-8_18 ◽

2022 ◽

pp. 279-289

Author(s):

Yunting Pu ◽

Federica Brandizzi

Keyword(s):

Arabidopsis Thaliana ◽

Unfolded Protein Response ◽

Proteomic Analysis ◽

Protein Preparation ◽

Unfolded Protein ◽

The Unfolded Protein Response ◽

Protein Response

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Proteomic Analysis of Glutathione S-Transferases of Arabidopsis thaliana Reveals Differential Salicylic Acid-Induced Expression of the Plant-Specific Phi and Tau Classes

Plant Molecular Biology ◽

10.1023/b:plan.0000028786.57439.b3 ◽

2004 ◽

Vol 54 (2) ◽

pp. 205-219 ◽

Cited By ~ 74

Author(s):

Pia G. Sappl ◽

Luis Oñate-Sánchez ◽

Karam B. Singh ◽

A. Harvey Millar

Keyword(s):

Arabidopsis Thaliana ◽

Salicylic Acid ◽

Proteomic Analysis ◽

Induced Expression ◽

Glutathione S Transferases

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Overexpression of chloroplast-targeted ferrochelatase 1 results in a genomes uncoupled chloroplast-to-nucleus retrograde signalling phenotype

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2019.0401 ◽

2020 ◽

Vol 375 (1801) ◽

pp. 20190401 ◽

Cited By ~ 2

Author(s):

Mike T. Page ◽

Tania Garcia-Becerra ◽

Alison G. Smith ◽

Matthew J. Terry

Keyword(s):

Gene Expression ◽

Feedback Inhibition ◽

Nuclear Gene ◽

Signalling Pathway ◽

Chloroplast Genes ◽

Nuclear Gene Expression ◽

Retrograde Signalling ◽

Genes Encoding ◽

Retrograde Signal

Chloroplast development requires communication between the progenitor plastids and the nucleus, where most of the genes encoding chloroplast proteins reside. Retrograde signals from the chloroplast to the nucleus control the expression of many of these genes, but the signalling pathway is poorly understood. Tetrapyrroles have been strongly implicated as mediators of this signal with the current hypothesis being that haem produced by the activity of ferrochelatase 1 (FC1) is required to promote nuclear gene expression. We have tested this hypothesis by overexpressing FC1 and specifically targeting it to either chloroplasts or mitochondria, two possible locations for this enzyme. Our results show that targeting of FC1 to chloroplasts results in increased expression of the nuclear-encoded chloroplast genes GUN4 , CA1 , HEMA1 , LHCB2.1, CHLH after treatment with Norflurazon (NF) and that this increase correlates to FC1 gene expression and haem production measured by feedback inhibition of protochlorophyllide synthesis. Targeting FC1 to mitochondria did not enhance the expression of nuclear-encoded chloroplast genes after NF treatment. The overexpression of FC1 also increased nuclear gene expression in the absence of NF treatment, demonstrating that this pathway is operational in the absence of a stress treatment. Our results therefore support the hypothesis that haem synthesis is a promotive chloroplast-to-nucleus retrograde signal. However, not all FC1 overexpression lines enhanced nuclear gene expression, suggesting there is still a lot we do not understand about the role of FC1 in this signalling pathway. This article is part of the theme issue ‘Retrograde signalling from endosymbiotic organelles’.

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