C172S Substitution in the Chloroplast-encoded Large Subunit Affects Stability and Stress-induced Turnover of Ribulose-1,5-bisphosphate Carboxylase/Oxygenase

Moreno, J.; Spreitzer, Robert J.

doi:10.1074/jbc.274.38.26789

Cited by 47 publications

(77 citation statements)

References 40 publications

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“…The Ats1A steady-state level of transcripts seemed also to be positively regulated by NaCl treatment, and negatively regulated by the treatment to PEG. This result is consistent with previous studies showing that the activity of the Rubisco enzyme is inhibited by an increase in the concentration of Pro residue (Sivakumar et al, 2000) and by a hyperosmotic treatment with mannitol (Moreno and Spreitzer, 1999). Conversely, NaCl has a modulating effect on the activity of the Rubisco (Sivakumar et al, 2000).…”

Section: Relative Expression Profile Of the 10 Atsk Genes Under Abiotsupporting

confidence: 82%

Expression Profiling of the Whole Arabidopsis Shaggy-Like Kinase Multigene Family by Real-Time Reverse Transcriptase-Polymerase Chain Reaction

et al. 2002

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The recent publication of the complete sequence of the Arabidopsis genome allowed us to identify and characterize the last two members of the SHAGGY-like kinase (AtSK) gene family. As a result, the study of the overall spatio-temporal organization of the whole AtSK family in Arabidopsis has become an achievable and necessary aim to understand the role of each SHAGGY-like kinase during plant development. An analysis of the transcript level of the 10 members of the family has been performed using the technique of real-time quantitative reverse transcriptase-polymerase chain reaction. Transcript levels in several organs, under different growth conditions, were analyzed. To calibrate the results obtained, a number of other genes, such as those coding for the two MAP3K⑀s and the two MAP4K␣s, as well as the stress response marker RD29A; the small subunit of the Rubisco photosynthetic enzyme Ats1A; the MEDEA chromatin remodeling factor; and the SCARECROW, ASYMMETRIC LEAVES 1, and SUPERMAN transcription factors all involved in key steps of plant development were used. The analysis of our data revealed that eight of the 10 genes of the AtSK family displayed a pseudo-constitutive expression pattern at the organ level. Conversely, AtSK13 responded to osmotic changes and saline treatment, whereas AtSK31 was flower specific and responded to osmotic changes and darkness.The SHAGGY/GSK3-like kinases are non-receptor Ser-Thr (S/T) kinases playing numerous roles (for review, see Kim and Kimmel, 2000). In animals, they are involved in the determination of cell destiny, resulting in the spatial organization of the body plan. In Drosophila melanogaster, a pool of several isoenzymes called SHAGGY and encoded by a single gene, is involved both in the definition of boundaries between the embryonic segments of the larvae (Siegfried et al., 1992), and in the development of the central and peripheral nervous system (Heitzler and Simpson, 1991). In the sea urchin embryo, the SHAGGY-like enzyme is involved in the definition of the animal/vegetal axis (Emily-Fenouil et al., 1998). In Xenopus laevis embryo, a deficiency for the activity of this kinase results in a defect of the dorso-ventral plan formation, leading to the formation of two heads (He et al., 1995). Finally, in mammals, two enzymes named GSK3␣ and GSK3␤ (for glycogensynthase kinase), encoded by two genes, are involved in the regulation of glycogen metabolism (Oreñ a et al., 2000), in the stability of the cytoskeleton (Zumbrunn et al., 2001), and in numerous other processes related to oncogenesis (Webster et al., 2001).In higher plants, the SHAGGY-like genes are present as small gene families. They have been characterized from a number of plant species (Pay et al., 1993; Tichtinsky et al., 1998; Jonak et al., 2000). Before the completion of the sequencing of the whole Arabidopsis genome, eight genes were known to belong to the SHAGGY-like gene family (AtSK; Jonak et al., 1995; Dornelas et al., 1998 Dornelas et al., , 1999 Tichtinsky et al., 1998). Recently, two additional ge...

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Section: Relative Expression Profile Of the 10 Atsk Genes Under Abiotsupporting

confidence: 82%

Expression Profiling of the Whole Arabidopsis Shaggy-Like Kinase Multigene Family by Real-Time Reverse Transcriptase-Polymerase Chain Reaction

et al. 2002

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“…Since the four mutants grew much like the wild type, it was evident that replacement of either cysteine residue did not affect synthesis and assembly of a functional enzyme crucial for photoautotrophic growth. A similar observation was reported for a C172S substitution in RuBisCO of C. reinhardtii (28). The prominent activity of RuBisCO in the C172A and C192A cyanobacterial mutants (Table 1), the ability of these mutants to grow photoautotrophically, and the lack of interaction between these residues and RuBP in RuBisCO crystals (29) indicate quite explicitly that neither Cys172 nor Cys192 is involved in catalysis, as was previously suggested (34).…”

Section: Discussionsupporting

confidence: 63%

Dual Role of Cysteine 172 in Redox Regulation of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase Activity and Degradation

et al. 2003

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Alkylation and oxidation of cysteine residues significantly decrease the catalytic activity and stimulate the degradation of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO). We analyzed the role of vicinal cysteine residues in redox regulation of RuBisCO from Synechocystis sp. strain PCC 6803. Cys172 and Cys192, which are adjacent to the catalytic site, and Cys247, which cross-links two large subunits, were replaced by alanine. Whereas all mutant cells (C172A, C192A, C172A-C192A, and C247A) and the wild type grew photoautotrophically at similar rates, the maximal photosynthesis rates of C172A mutants decreased 10 to 20% as a result of 40 to 60% declines in RuBisCO turnover number. Replacement of Cys172, but not replacement of Cys192, prominently decreased the effect of cysteine alkylation or oxidation on RuBisCO. Oxidants that react with vicinal thiols had a less inhibitory effect on the activity of either the C172A or C192A enzyme variants, suggesting that a disulfide bond was formed upon oxidation. Thiol oxidation induced RuBisCO dissociation into subunits. This effect was either reduced in the C172A and C192A mutant enzymes or eliminated by carboxypentitol bisphosphate (CPBP) binding to the activated enzyme form. The CPBP effect presumably resulted from a conformational change in the carbamylated CPBP-bound enzyme, as implied from an alteration in the electrophoretic mobility. Stress conditions, provoked by nitrate deprivation, decreased the RuBisCO contents and activities in the wild type and in the C192A and C247A mutants but not in the C172A and C172A-C192A mutants. These results suggest that although Cys172 does not participate in catalysis, it plays a role in redox regulation of RuBisCO activity and degradation.The universal CO 2 -fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) (EC 4.1.1.39) catalyzes the primary reactions of photosynthesis and photorespiration by carboxylation and oxygenation of ribulose-1,5-bisphosphate (RuBP), respectively. The RuBisCO from cyanobacteria, algae (excluding some dinoflagellates), and higher plants is a hexadecamer consisting of eight large and eight small subunits organized as four protomers, each of which has two active sites at the interface between each large subunit pair (3,4,29). In the RuBisCO of many species, an intradimeric disulfide bond is formed under oxidizing conditions between the Cys247 residues of a large subunit pair (30). A sequence comparison of more than 500 RuBisCO genes revealed high levels of similarity, particularly among large subunits, and residues that form the catalytic site are entirely conserved (21). As a result of the enzymatic inefficiency manifested by a low catalytic turnover number (1.5 to 12 carboxylations s Ϫ1 catalytic site Ϫ1 [5,24]), as well as a low affinity for substrate CO 2 and competition between the CO 2 and O 2 substrates, it is accepted that the activity of RuBisCO limits both photosynthesis and photorespiration under various ecophysiological conditions (40).Due to the pivotal role of RuB...

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“…The method employed to determine glutathionylation sites appears to be reliable, because many of the sites identified correspond to cysteines known to undergo glutathionylation or oxidative modifications such as Cys 217 in human actin (58), Cys 385 in porcine aconitase (59), Cys 98 in yeast PGK (60), Cys 247 in Chlamydomonas isocitrate lyase (20,23), and Cys 172 in Chlamydomonas Rubisco (61). A search for motifs in the primary sequence surrounding the 56 sites identified did not reveal any consensus sequence.…”

Section: Discussionmentioning

confidence: 99%

Glutathionylation in the Photosynthetic Model Organism Chlamydomonas reinhardtii: A Proteomic Survey

Zaffagnini

Bedhomme

Groni

et al. 2012

Molecular & Cellular Proteomics

136

164

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Protein glutathionylation is a redox post-translational modification occurring under oxidative stress conditions and playing a major role in cell regulation and signaling. This modification has been mainly studied in nonphotosynthetic organisms, whereas much less is known in photosynthetic organisms despite their important exposure to oxidative stress caused by changes in environmental conditions. We report a large scale proteomic analysis using biotinylated glutathione and streptavidin affinity chromatography that allowed identification of 225 glutathionylated proteins in the eukaryotic unicellular green alga Chlamydomonas reinhardtii. Moreover, 56 sites of glutathionylation were also identified after peptide affinity purification and tandem mass spectrometry. The targets identified belong to a wide range of biological processes and pathways, among which the Calvin-Benson cycle appears to be a major target. The glutathionylation of four enzymes of this cycle, phosphoribulokinase, glyceraldehyde-3-phosphate dehydrogenase, ribose-5-phosphate isomerase, and phosphoglycerate kinase was confirmed by Western blot and activity measurements. The results suggest that glutathionylation could constitute a major mechanism of regulation of the Calvin-Benson cycle under oxidative stress conditions. Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.014142, 1-15, 2012.Protein post-translational modifications (PTMs) 1 play a pivotal role in cellular signaling (1). Recently, redox PTMs have emerged as important mechanisms of signaling and regulation in all organisms. Our increasing understanding of the molecular mechanism of cell signaling has revealed that reactive oxygen species (ROS) and reactive nitrogen species (RNS) act as signaling molecules to transfer extracellular or intracellular information and elicit specific responses. ROS and RNS have generally been considered to be toxic molecules that have to be continuously scavenged and efficiently detoxified. Plant cells exhibit a remarkable ability to cope with high rates of ROS/RNS production as a result of a complex scavenging system that includes either antioxidant molecules or enzymes (2). In photosynthetic organisms, ROS and RNS are continuously produced during normal aerobic metabolism but are also produced transiently in response to various types of endogenous or exogenous signals, such as biotic and abiotic stresses. This production activates specific signaling pathways, resulting in transcriptional, post-transcriptional and post-translational responses that will, in fine, allow adaptation to new environmental conditions. These past decades, redox modifications have emerged as central mechanisms in these processes, at the interface between ROS/RNS and the adaptative responses to environmental changes. ROS/RNS signaling operates mainly through a set of PTMs of thiol residues on proteins (3). Indeed, cysteine residues can undergo different states of oxidation such as sulfenic, sulfinic, and sulfonic acids in addition to protein disulfide bridges (intra-or intermolec...

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C172S Substitution in the Chloroplast-encoded Large Subunit Affects Stability and Stress-induced Turnover of Ribulose-1,5-bisphosphate Carboxylase/Oxygenase

Cited by 47 publications

References 40 publications

Expression Profiling of the Whole Arabidopsis Shaggy-Like Kinase Multigene Family by Real-Time Reverse Transcriptase-Polymerase Chain Reaction

Expression Profiling of the Whole Arabidopsis Shaggy-Like Kinase Multigene Family by Real-Time Reverse Transcriptase-Polymerase Chain Reaction

Dual Role of Cysteine 172 in Redox Regulation of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase Activity and Degradation

Glutathionylation in the Photosynthetic Model Organism Chlamydomonas reinhardtii: A Proteomic Survey

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