N-terminus plus linker domain of Mg-chelatase D subunit is essential for Mg-chelatase activity in Oryza sativa

Luo, Sha; Luo, Tao; Liu, Yinan; Li, Zunwen; Fan, Shiliang; Wu, Caijun

doi:10.1016/j.bbrc.2018.02.146

Cited by 7 publications

(12 citation statements)

References 22 publications

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“…Thus, mass spectrometry and MST show that the C-terminal integrin I domain of ChlD is the major determinant of binding to ChlH. In the Luo et al study [15], where recombinant proteins from rice are used, their results suggest that only the AAA + and linker domains are required for protein activity, results that directly contradict this work, previous studies from our laboratory [11,13] and that of other groups [14]. While it is possible that higher plant enzymes have evolved different interaction sites between the ChlD and ChlH proteins, the proposals of Luo et al [15] require biophysical characterisation of the protein–protein interactions of the rice proteins.…”

Section: Resultsmentioning

confidence: 99%

“…While it is known that ChlD and ChlI form a complex [8,12], the connection between ChlID and ChlH was less well defined. Previous studies have described the importance of the polyproline linker and AAA + domain in ChlD to the activity of MgCH [12,15,41,42], but there has been limited description of the role of the C-terminal integrin I domain of ChlD, other than the work of Axelsson et al [14] which revealed the importance of the MIDAS residues for chelatase activity. Our work has provided a biochemical and kinetic counterpart to the results of the recent study by Luo et al [15], which used yeast two-hybrid analysis to show that ChlD interacts with ChlH, although our work proposes a different location for this protein–protein interaction.…”

Section: Resultsmentioning

confidence: 99%

“…Previous studies have described the importance of the polyproline linker and AAA + domain in ChlD to the activity of MgCH [12,15,41,42], but there has been limited description of the role of the C-terminal integrin I domain of ChlD, other than the work of Axelsson et al [14] which revealed the importance of the MIDAS residues for chelatase activity. Our work has provided a biochemical and kinetic counterpart to the results of the recent study by Luo et al [15], which used yeast two-hybrid analysis to show that ChlD interacts with ChlH, although our work proposes a different location for this protein–protein interaction. We have previously shown that C- and N-terminal truncations of ChlD from Synechocystis and Thermosynechoccus elongatus are not able to produce active chelatases [11,13], and that the N-terminal AAA + domain of ChlD is primarily responsible for interaction with ChlI [11].…”

Section: Resultsmentioning

confidence: 99%

“…PCC 6803 (hereafter Synechocystis ) MgCH, although not in the Thermosynechococcus elongatus enzyme [13]. Previous work has shown the importance of a fully intact ChlD (or bacterial homologue, BchD) protein for maintaining chelatase activity [11,13,14], although recent work published by Luo et al [15] suggests only the N-terminal AAA + and polyproline linker domains are required for activity.…”

Section: Introductionmentioning

confidence: 99%

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The ChlD subunit links the motor and porphyrin binding subunits of magnesium chelatase

Farmer

Brindley

Hitchcock

et al. 2019

Biochemical Journal

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Magnesium chelatase initiates chlorophyll biosynthesis, catalysing the MgATP2−-dependent insertion of a Mg2+ ion into protoporphyrin IX. The catalytic core of this large enzyme complex consists of three subunits: Bch/ChlI, Bch/ChlD and Bch/ChlH (in bacteriochlorophyll and chlorophyll producing species, respectively). The D and I subunits are members of the AAA+ (ATPases associated with various cellular activities) superfamily of enzymes, and they form a complex that binds to H, the site of metal ion insertion. In order to investigate the physical coupling between ChlID and ChlH in vivo and in vitro, ChlD was FLAG-tagged in the cyanobacterium Synechocystis sp. PCC 6803 and co-immunoprecipitation experiments showed interactions with both ChlI and ChlH. Co-production of recombinant ChlD and ChlH in Escherichia coli yielded a ChlDH complex. Quantitative analysis using microscale thermophoresis showed magnesium-dependent binding (Kd 331 ± 58 nM) between ChlD and H. The physical basis for a ChlD–H interaction was investigated using chemical cross-linking coupled with mass spectrometry (XL–MS), together with modifications that either truncate ChlD or modify single residues. We found that the C-terminal integrin I domain of ChlD governs association with ChlH, the Mg2+ dependence of which also mediates the cooperative response of the Synechocystis chelatase to magnesium. The interaction site between the AAA+ motor and the chelatase domain of magnesium chelatase will be essential for understanding how free energy from the hydrolysis of ATP on the AAA+ ChlI subunit is transmitted via the bridging subunit ChlD to the active site on ChlH.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The ChlD subunit links the motor and porphyrin binding subunits of magnesium chelatase

Farmer

Brindley

Hitchcock

et al. 2019

Biochemical Journal

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“…ChlH is a porphyrin-binding subunit and a key enzyme in chlorophyll synthesis [25]. Mg-chelatase makes protoporphyrin IX participate in chlorophyll biosynthesis, and its activity decreases with decreasing ChlH transcription levels [26]. In this study, ChlH and other genes involved in chlorophyll synthesis (HemA, HemC, HemE) were downregulated.…”

Section: Discussionmentioning

confidence: 60%

Physiological adaptation and transcriptome changes in the plateau plant common vetch (Vicia sativa L.) in response to the plain environment

Kong

et al. 2021

Preprint

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Background Light and temperature are among the most important factors in plant growth. The annual forage variety common vetch was grown under the special environment of high light intensity and low temperature on the Qinghai-Tibet Plateau. In the seedling stage, the Qinghai-Tibet Plateau adapted variety common vetch was airlifted to the inland plain. Its response to the plain environment and its regulatory mechanism are not clear. Results The samples collected the day before transportation were used as a control. After being transported back to the plain, the morphological structure and physiological indexes were determined on days one, three, five and seven. Compared with the control group, the chlorophyll content in the experimental group changed significantly. Chlorophyll fluorescence increased significantly on the fifth day. The number of stomatal openings decreased significantly on the first day and then increased gradually and exceeded that of the control. Compared with the control sample, the intercellular space of the experimental sample was larger and then did not return to the control level. The physiological indexes gradually returned to the control level on the 7th day. Transcriptome analysis showed that 3251 genes were upregulated and 3317 genes were downregulated on the third day, while 1359 genes were upregulated and 1648 genes were downregulated on the seventh day. These differentially expressed genes were significantly enriched in photosynthesis, photosynthetic antenna protein synthesis, carbon dioxide fixation and chlorophyll synthesis pathways, and almost all of the genes involved in these pathways were downregulated. In addition, MYB, NAC, AP2-EREBP and the Orphans family of transcription factors (TFs) regulating the light response were more abundant on day 3 and day 7 than in the control group. Conclusion The physiological and gene expression levels of common vetch transported to the plain environment were analyzed. To reveal the adaptation mechanism of common vetch to plain environments, the key metabolic pathways of differential gene enrichment were analyzed. These findings are helpful for introducing plateau suitable varieties into plain environments.

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Mapping and Functional Analysis of LE Gene in a Lethal Etiolated Rice Mutant at Seedling Stage

Xiaodong,

Xiaobo,

Zhonghao

et al. 2023

Rice Science

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N-terminus plus linker domain of Mg-chelatase D subunit is essential for Mg-chelatase activity in Oryza sativa

Cited by 7 publications

References 22 publications

The ChlD subunit links the motor and porphyrin binding subunits of magnesium chelatase

The ChlD subunit links the motor and porphyrin binding subunits of magnesium chelatase

Physiological adaptation and transcriptome changes in the plateau plant common vetch (Vicia sativa L.) in response to the plain environment

Mapping and Functional Analysis of LE Gene in a Lethal Etiolated Rice Mutant at Seedling Stage

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