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Title: Identification of the plant ribokinase and discovery of a role for Arabidopsis ribokinase in nucleoside metabolism

Abstract

Ribose can be used for energy or as a component of several important biomolecules, but for it to be used in either capacity it must first be phosphorylated by ribokinase (RBSK). RBSK proteins are part of the phosphofructokinase-B (pfkB) family of carbohydrate kinases. Sequence comparisons of pfkB proteins from the model plant Arabidopsis thaliana with the human and Escherichia coli RBSK identified a single candidate RBSK, At1g17160 (AtRBSK). AtRBSK is more similar to predicted RBSKs from other plant species and known mammalian and prokaryotic RBSK than to all other PfkB proteins in Arabidopsis. AtRBSK contains a predicted chloroplast transit peptide, and we confirmed plastid localization using AtRBSK fused to YFP. Structure prediction software verified that the AtRBSK sequence mapped onto a known RBSK structure. Kinetic parameters of purified recombinant AtRBSK were determined to be K mribose = 150 μm ± 17 μm, K mATP = 45 μm ± 5.6 μm, and k cat = 2.0 s ₋1. Substrate inhibition was observed for AtRBSK (K iATP = 2.44 mm ± 0.36 mm), as has been demonstrated for other RBSK proteins. Ribose accumulated in Arabidopsis plants lacking AtRBSK. Such plants grew normally unless media was supplemented with ribose, which led to chlorosismore » and growth inhibition. Both chlorosis and ribose accumulation were abolished upon the introduction of a transgene expressing AtRBSK-MYC, demonstrating that the loss of protein is responsible for ribose hypersensitivity. Lastly, ribose accumulation in plants lacking AtRBSK was reduced in plants also deficient in the nucleoside ribohydrolase NSH1, linking AtRBSK activity to nucleoside metabolism.« less

Authors:
 [1];  [1];  [1]; ORCiD logo [1]
  1. Univ. of California, Davis, CA (United States). Dept. of Molecular and Cellular Biology
Publication Date:
Research Org.:
Univ. of California, Davis, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1333861
Grant/Contract Number:  
SC0002175
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 291; Journal Issue: 43; Journal ID: ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular Biology
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; carbohydrate metabolism; carbohydrate kinase; kinase; ribokinase

Citation Formats

Riggs, John W., Rockwell, Nathan C., Cavales, Philip C., and Callis, Judy. Identification of the plant ribokinase and discovery of a role for Arabidopsis ribokinase in nucleoside metabolism. United States: N. p., 2016. Web. doi:10.1074/jbc.M116.754689.
Riggs, John W., Rockwell, Nathan C., Cavales, Philip C., & Callis, Judy. Identification of the plant ribokinase and discovery of a role for Arabidopsis ribokinase in nucleoside metabolism. United States. doi:10.1074/jbc.M116.754689.
Riggs, John W., Rockwell, Nathan C., Cavales, Philip C., and Callis, Judy. Tue . "Identification of the plant ribokinase and discovery of a role for Arabidopsis ribokinase in nucleoside metabolism". United States. doi:10.1074/jbc.M116.754689. https://www.osti.gov/servlets/purl/1333861.
@article{osti_1333861,
title = {Identification of the plant ribokinase and discovery of a role for Arabidopsis ribokinase in nucleoside metabolism},
author = {Riggs, John W. and Rockwell, Nathan C. and Cavales, Philip C. and Callis, Judy},
abstractNote = {Ribose can be used for energy or as a component of several important biomolecules, but for it to be used in either capacity it must first be phosphorylated by ribokinase (RBSK). RBSK proteins are part of the phosphofructokinase-B (pfkB) family of carbohydrate kinases. Sequence comparisons of pfkB proteins from the model plant Arabidopsis thaliana with the human and Escherichia coli RBSK identified a single candidate RBSK, At1g17160 (AtRBSK). AtRBSK is more similar to predicted RBSKs from other plant species and known mammalian and prokaryotic RBSK than to all other PfkB proteins in Arabidopsis. AtRBSK contains a predicted chloroplast transit peptide, and we confirmed plastid localization using AtRBSK fused to YFP. Structure prediction software verified that the AtRBSK sequence mapped onto a known RBSK structure. Kinetic parameters of purified recombinant AtRBSK were determined to be Kmribose = 150 μm ± 17 μm, KmATP = 45 μm ± 5.6 μm, and kcat = 2.0 s₋1. Substrate inhibition was observed for AtRBSK (KiATP = 2.44 mm ± 0.36 mm), as has been demonstrated for other RBSK proteins. Ribose accumulated in Arabidopsis plants lacking AtRBSK. Such plants grew normally unless media was supplemented with ribose, which led to chlorosis and growth inhibition. Both chlorosis and ribose accumulation were abolished upon the introduction of a transgene expressing AtRBSK-MYC, demonstrating that the loss of protein is responsible for ribose hypersensitivity. Lastly, ribose accumulation in plants lacking AtRBSK was reduced in plants also deficient in the nucleoside ribohydrolase NSH1, linking AtRBSK activity to nucleoside metabolism.},
doi = {10.1074/jbc.M116.754689},
journal = {Journal of Biological Chemistry},
number = 43,
volume = 291,
place = {United States},
year = {2016},
month = {9}
}

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