Purification and biochemical characterization of Arabidopsis At-NEET, an ancient iron-sulfur protein, reveals a conserved cleavage motif for subcellular localization

Su, Liwen; Chang, Shu Heng; Li, Mengying; Huang, Heng-Yi; Jane, Wann‐Neng; Yang, Jun-Yi

doi:10.1016/j.plantsci.2013.09.001

Cited by 20 publications

(21 citation statements)

References 33 publications

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“…AtNEET has been localized previously in both mitochondria and chloroplasts, and has been found to be a key regulator in plant development, senescence, iron metabolism and ROS homeostasis (Nechushtai et al ., ). Contradictory results on its subcellular localization have also been reported, as a recent study concluded that AtNEET translocates in the chloroplast, but not in mitochondria (Su et al ., ). With NtNEET, we found that the protein was localized only in the chloroplast.…”

Section: Discussionmentioning

confidence: 97%

“…The transport of proteins to mitochondria and chloroplasts can be achieved through N‐terminal transit peptides. Accordingly, AtNEET and NtNEET contain the proposed N‐terminal chloroplast transit peptide cleavage motif V‐R/K‐A‐E (Su et al ., ). However, this N‐terminal region presents most of the differences observed in their sequences.…”

Section: Discussionmentioning

confidence: 97%

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The coat protein of Alfalfa mosaic virus interacts and interferes with the transcriptional activity of the bHLH transcription factor ILR3 promoting salicylic acid‐dependent defence signalling response

Aparicio

Pallás

2016

Molecular Plant Pathology

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During virus infection, specific viral component-host factor interaction elicits the transcriptional reprogramming of diverse cellular pathways. Alfalfa mosaic virus (AMV) can establish a compatible interaction in tobacco and Arabidopsis hosts. We show that the coat protein (CP) of AMV interacts directly with transcription factor (TF) ILR3 of both species. ILR3 is a basic helix-loop-helix (bHLH) family member of TFs, previously proposed to participate in diverse metabolic pathways. ILR3 has been shown to regulate NEET in Arabidopsis, a critical protein in plant development, senescence, iron metabolism and reactive oxygen species (ROS) homeostasis. We show that the AMV CP-ILR3 interaction causes a fraction of this TF to relocate from the nucleus to the nucleolus. ROS, pathogenesis-related protein 1 (PR1) mRNAs, salicylic acid (SA) and jasmonic acid (JA) contents are increased in healthy Arabidopsis loss-of-function ILR3 mutant (ilr3.2) plants, which implicates ILR3 in the regulation of plant defence responses. In AMV-infected wild-type (wt) plants, NEET expression is reduced slightly, but is induced significantly in ilr3.2 mutant plants. Furthermore, the accumulation of SA and JA is induced in Arabidopsis wt-infected plants. AMV infection in ilr3.2 plants increases JA by over 10-fold, and SA is reduced significantly, indicating an antagonist crosstalk effect. The accumulation levels of viral RNAs are decreased significantly in ilr3.2 mutants, but the virus can still systemically invade the plant. The AMV CP-ILR3 interaction may down-regulate a host factor, NEET, leading to the activation of plant hormone responses to obtain a hormonal equilibrium state, where infection remains at a level that does not affect plant viability.

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 97%

The coat protein of Alfalfa mosaic virus interacts and interferes with the transcriptional activity of the bHLH transcription factor ILR3 promoting salicylic acid‐dependent defence signalling response

Aparicio

Pallás

2016

Molecular Plant Pathology

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“…has the additional advantage that it contains only a single homologous NEET gene, At5g51720 (At-NEET) [31]. At-NEET was found to localize to the chloroplast and to the mitochondria [31,59]. Phenotypic characterization of At-NEET knockdown plants revealed a key role for this protein in plant development, senescence, reactive oxygen homeostasis, and Fe metabolism (Fig.…”

Section: Neet Proteins Are Involved In Cellular and Whole Organism Irmentioning

confidence: 99%

Structure–function analysis of NEET proteins uncovers their role as key regulators of iron and ROS homeostasis in health and disease

Tamir

Paddock

Darash-Yahana-Baram

et al. 2015

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

137

230

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A novel family of 2Fe-2S proteins, the NEET family, was discovered during the last decade in numerous organisms, including archea, bacteria, algae, plant and human; suggesting an evolutionary-conserved function, potentially mediated by their CDGSH Iron-Sulfur Domain. In human, three NEET members encoded by the CISD1-3 genes were identified. The structures of CISD1 (mitoNEET, mNT), CISD2 (NAF-1), and the plant At-NEET uncovered a homodimer with a unique "NEET fold", as well as two distinct domains: a beta-cap and a 2Fe-2S cluster-binding domain. The 2Fe-2S clusters of NEET proteins were found to be coordinated by a novel 3Cys:1His structure that is relatively labile compared to other 2Fe-2S proteins and is the reason of the NEETs' clusters could be transferred to apo-acceptor protein(s) or mitochondria. Positioned at the protein surface, the NEET's 2Fe-2S's coordinating His is exposed to protonation upon changes in its environment, potentially suggesting a sensing function for this residue. Studies in different model systems demonstrated a role for NAF-1 and mNT in the regulation of cellular iron, calcium and ROS homeostasis, and uncovered a key role for NEET proteins in critical processes, such as cancer cell proliferation and tumor growth, lipid and glucose homeostasis in obesity and diabetes, control of autophagy, longevity in mice, and senescence in plants. Abnormal regulation of NEET proteins was consequently found to result in multiple health conditions, and aberrant splicing of NAF-1 was found to be a causative of the neurological genetic disorder Wolfram Syndrome 2. Here we review the discovery of NEET proteins, their structural, biochemical and biophysical characterization, and their most recent structure-function analyses. We additionally highlight future avenues of research focused on NEET proteins and propose an essential role for NEETs in health and disease. This article is part of a Special Issue entitled: Fe/S proteins: Analysis, structure, function, biogenesis and diseases.

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“…1), resides both in chloroplast and mitochondria. At-NEET has a key role in plant development, senescence, reactive oxygen homeostasis, iron metabolism and homeostasis in different cells [30–32]. At-NEET encoded by the (At5g51720) gene shows 50 and 57% similarity to mNT and NAF-1, respectively, while its [2Fe-2S] binding domain has sequence identity to mNT and NAF-1 of about 75 and 88%, respectively [30, 32].…”

Section: Prefacementioning

confidence: 99%

The unique fold and lability of the [2Fe-2S] clusters of NEET proteins mediate their key functions in health and disease

et al. 2018

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NEET proteins comprise a new class of [2Fe-2S] cluster proteins. In human, three genes encode for NEET proteins: cisd1 encodes mitoNEET (mNT), cisd2 encodes the Nutrient-deprivation autophagy factor-1 (NAF-1) and cisd3 encodes MiNT (Miner2). These recently discovered proteins play key roles in many processes related to normal metabolism and disease. Indeed, NEET proteins are involved in iron, Fe-S, and reactive oxygen homeostasis in cells and play an important role in regulating apoptosis and autophagy. mNT and NAF-1 are homodimeric and reside on the outer mitochondrial membrane. NAF-1 also resides in the membranes of the ER associated mitochondrial membranes (MAM) and the ER. MiNT is a monomer with distinct asymmetry in the molecular surfaces surrounding the clusters. Unlike its paralogs mNT and NAF-1, it resides within the mitochondria. NAF-1 and mNT share similar backbone folds to the plant homodimeric NEET protein (At-NEET), while MiNT’s backbone fold resembles a bacterial MiNT protein. Despite the variation of amino acid composition among these proteins, all NEET proteins retained their unique CDGSH domain harboring their unique 3Cys:1His [2Fe-2S] cluster coordination through evolution. The coordinating exposed His was shown to convey the lability to the NEET proteins’ [2Fe-2S] clusters. In this minireview, we discuss the NEET fold and its structural elements. Special attention is given to the unique lability of the NEETs’ [2Fe-2S] cluster and the implication of the latter to the NEET proteins’ cellular and systemic function in health and disease.Graphical abstract

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Purification and biochemical characterization of Arabidopsis At-NEET, an ancient iron-sulfur protein, reveals a conserved cleavage motif for subcellular localization

Cited by 20 publications

References 33 publications

The coat protein of Alfalfa mosaic virus interacts and interferes with the transcriptional activity of the bHLH transcription factor ILR3 promoting salicylic acid‐dependent defence signalling response

The coat protein of Alfalfa mosaic virus interacts and interferes with the transcriptional activity of the bHLH transcription factor ILR3 promoting salicylic acid‐dependent defence signalling response

Structure–function analysis of NEET proteins uncovers their role as key regulators of iron and ROS homeostasis in health and disease

The unique fold and lability of the [2Fe-2S] clusters of NEET proteins mediate their key functions in health and disease

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