Proteomic Approach to Identify Novel Mitochondrial Proteins in Arabidopsis

Kruft, Volker; Eubel, Holger; Jänsch, Lothar; Werhahn, Wolf; Braun, Hans‐Peter

doi:10.1104/pp.010474

Cited by 335 publications

(183 citation statements)

References 67 publications

(23 reference statements)

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“…The 80S ribosomes were obtained from 500 g Arabidopsis thaliana rosette leaves according to the protocol established by Kruft et al (2001), with few modifications. The entire procedure was performed at 4°C in the presence of the protease inhibitor cocktail complex (Roche, Mannheim, Germany).…”

Section: Preparation Of the 80s Ribosomesmentioning

confidence: 99%

“…In this study, the preparation of the ribosomal particles from the A. thaliana whole leaf was performed using the protocol developed by Kruft et al (2001), which is among one of the most common protocol used for the preparation of intact 80S plant ribosomes. The method employs a series of differential centrifugations, which lead to the preparation of a crude ribosomal fraction.…”

Section: Separation Of the Crude And 80s Ribosomal Proteins Using 2-dementioning

confidence: 99%

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High heterogeneity within the ribosomal proteins of the Arabidopsis thaliana 80S ribosome

et al. 2005

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Proteomic studies have addressed the composition of plant chloroplast ribosomes and 70S ribosomes from the unicellular organism Chlamydomonas reinhardtii, but comprehensive characterization of cytoplasmic 80S ribosomes from higher plants has been lacking. We have used two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) to analyse the cytoplasmic 80S ribosomes from the model flowering plant Arabidopsis thaliana. Of the 80 ribosomal protein families predicted to comprise the cytoplasmic 80S ribosome, we have confirmed the presence of 61; specifically, 27 (84%) of the small 40S subunit and 34 (71%) of the large 60S subunit. Nearly half (45%) of the ribosomal proteins identified are represented by two or more distinct spots in the 2-DE gel indicating that these proteins are either post-translationally modified or present as different isoforms. Consistently, MS-based protein identification revealed that at least one-third (34%) of the identified ribosomal protein families showed expression of two or more family members. In addition, we have identified a number of non-ribosomal proteins that co-migrate with the plant 80S ribosomes during gradient centrifugation suggesting their possible association with the 80S ribosomes. Among them, RACK1 has recently been proposed to be a ribosome-associated protein that promotes efficient translation in yeast. The study, thus provides the basis for further investigation into the function of the other identified non-ribosomal proteins as well as the biological meaning of the various ribosomal protein isoforms.

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Section: Preparation Of the 80s Ribosomesmentioning

confidence: 99%

Section: Separation Of the Crude And 80s Ribosomal Proteins Using 2-dementioning

confidence: 99%

High heterogeneity within the ribosomal proteins of the Arabidopsis thaliana 80S ribosome

et al. 2005

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“…Assignment of subcellular location to the encoded proteins was based on experimental evidence (Kruft et al, 2001;Friso et al, 2004;Heazlewood et al, 2004;Kleffmann et al, 2004) or on stringent computational prediction of subcellular targeting (Richly et al, 2003b;Richly and Leister, 2004). A list of all 1590 genes contained in the 23 regulons, as well as their assignment to functional classes and the subcellular location of the encoded proteins, is provided as Supplementary Material 2.…”

Section: Functional Classification and Assignment Of Subcellular Locamentioning

confidence: 99%

“…Assignment was based on experimental evidence (Kruft et al, 2001;Friso et al, 2004;Heazlewood et al, 2004;Kleffmann et al, 2004) or on stringent computational prediction of subcellular targeting (Richly et al, 2003b;Richly and Leister, 2004). Because the location of only a minority of chloroplast and mitochondrial proteins has been reliably experimentally determined, and because the computational prediction applied was of high specificity but relatively low sensitivity (Richly et al, 2003b;Richly and Leister, 2004), our classification underestimates the number of proteins located in chloroplasts or mitochondria (it follows that in Fig.…”

Section: Subcellular Protein Location Associated To Regulonsmentioning

confidence: 99%

Analysis of 101 nuclear transcriptomes reveals 23 distinct regulons and their relationship to metabolism, chromosomal gene distribution and co-ordination of nuclear and plastid gene expression

Biehl¹,

Richly

Noutsos

et al. 2005

Gene

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Post-endosymbiotic evolution of the proto-chloroplast was characterized by gene transfer to the nucleus. Hence, most chloroplast proteins are nuclear-encoded and the regulation of chloroplast functions includes nuclear transcriptional control. The expression profiles of 3292 nuclear Arabidopsis genes, most of them encoding chloroplast proteins, were determined from 101 different conditions and have been deposited at the GEO database (http://www.ncbi.nlm.nih.gov/geo/) under GSE1160-GSE1260. The 1590 most-regulated genes fell into 23 distinct groups of co-regulated genes (regulons). Genes of some regulons are not evenly distributed among the five Arabidopsis chromosomes and pairs of adjacent, co-expressed genes exist. Except regulons 1 and 2, regulons are heterogeneous and consist of genes coding for proteins with different subcellular locations or contributing to several biochemical functions. This implies that different organelles and/or metabolic pathways are co-ordinated at the nuclear transcriptional level, and a prototype for this is regulon 12 which contains genes with functions in amino acid and carbohydrate metabolism, as well as genes associated with transport or transcription. The co-expression of nuclear genes coding for subunits of the photosystems or encoding proteins involved in the transcription/translation of plastome genes (particularly ribosome polypeptides) (regulons 1 and 2, respectively) implies the existence of a novel mechanism that coordinates plastid and nuclear gene expression and involves nuclear control of plastid ribosome abundance. The co-regulation of genes for photosystem and plastid ribosome proteins escapes a previously described general control of nuclear chloroplast proteins imposed by a transcriptional master switch, highlighting a mode of transcriptional regulation of photosynthesis which is different compared to other chloroplast functions. From the evolutionary standpoint, the results provided indicate that functional integration of the proto-chloroplast into the eukaryotic cell was associated with the establishment of different layers of nuclear transcriptional control. D

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“…Six Prx II genes have been identified in Arabidopsis [6,11], one of them seems to be a pseudogene; the others are expressed in leaves. Three Prxs localized to the cytosol (AtPrx II B, C, and D), one to the chloroplast (AtPrx II E), and one to the mitochondria (AtPrx II F) [6,13,14].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Characterization of a Type II Peroxiredoxin Gene from Panax ginseng C. A. Meyer

Kim¹,

Lee²,

Lee³

et al. 2010

Journal of Ginseng Research

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A peroxiredoxin cDNA (PgPrx) was isolated and characterized from the leaves of Panax ginseng. The cDNA is 716 nucleotides long and has an open reading frame of 489 base pairs with a deduced amino acid sequence of 162 residues. The calculated molecular mass of the mature protein is approximately 17.4 kDa with a predicted isoelectric point of 5.37. A GenBank BlastX search revealed that the deduced amino acid sequence of PgPrx shares a high degree homology with type II peroxiredoxin (Prx) proteins in other plants. The PgPrx gene was highly expressed in leaves, and expressed at a low level in the stem. To analyze the gene expression of PgPrx in response to various abiotic stresses, we utilized real-time quantitative RT-PCR. Our results reveal that PgPrx expression is induced by ultraviolet irradiation, low temperature, and salt. The induction of PgPrx in response to abiotic stimuli suggests that ginseng Prx may function to protect the host against environmental stresses.

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Proteomic Approach to Identify Novel Mitochondrial Proteins in Arabidopsis

Cited by 335 publications

References 67 publications

High heterogeneity within the ribosomal proteins of the Arabidopsis thaliana 80S ribosome

High heterogeneity within the ribosomal proteins of the Arabidopsis thaliana 80S ribosome

Analysis of 101 nuclear transcriptomes reveals 23 distinct regulons and their relationship to metabolism, chromosomal gene distribution and co-ordination of nuclear and plastid gene expression

Isolation and Characterization of a Type II Peroxiredoxin Gene from Panax ginseng C. A. Meyer

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