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The role of HEXOKINASE1 in Arabidopsis leaf growth

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Here, we used a hxk1 mutant in the Col-0 background. We demonstrated that HXK1 regulates cell proliferation and expansion early during leaf development, and that HXK1 is involved in sucrose-induced leaf growth stimulation independent of GPT2. Furthermore, we identified KINγ as a novel HXK1-interacting protein.

Abstract

In the last decade, extensive efforts have been made to unravel the underlying mechanisms of plant growth control through sugar availability. Signaling by the conserved glucose sensor HEXOKINASE1 (HXK1) has been shown to exert both growth-promoting and growth-inhibitory effects depending on the sugar levels, the environmental conditions and the plant species. Here, we used a hxk1 mutant in the Col-0 background to investigate the role of HXK1 during leaf growth in more detail and show that it is affected in both cell proliferation and cell expansion early during leaf development. Furthermore, the hxk1 mutant is less sensitive to sucrose-induced cell proliferation with no significant increase in final leaf growth after transfer to sucrose. Early during leaf development, transfer to sucrose stimulates expression of GLUCOSE-6-PHOSPHATE/PHOSPHATE TRANSPORTER2 (GPT2) and represses chloroplast differentiation. However, in the hxk1 mutant GPT2 expression was still upregulated by transfer to sucrose although chloroplast differentiation was not affected, suggesting that GPT2 is not involved in HXK1-dependent regulation of leaf growth. Finally, using tandem affinity purification of protein complexes from cell cultures, we identified KINγ, a protein containing four cystathionine β-synthase domains, as an interacting protein of HXK1.

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References

  • Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P, Stevenson DK, Zimmerman J, Barajas P, Cheuk R, Gadrinab C, Heller C, Jeske A, Koesema E, Meyers CC, Parker H, Prednis L, Ansari Y, Choy N, Deen H, Geralt M, Hazari N, Hom E, Karnes M, Mulholland C, Ndubaku R, Schmidt I, Guzman P, Aguilar-Henonin L, Schmid M, Weigel D, Carter DE, Marchand T, Risseeuw E, Brogden D, Zeko A, Crosby WL, Berry CC, Ecker JR (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657

    Article  PubMed  Google Scholar 

  • Andriankaja M, Dhondt S, De Bodt S, Vanhaeren H, Coppens F, De Milde L, Mühlenbock P, Skirycz A, Gonzalez N, Beemster GTS, Inzé D (2012) Exit from proliferation during leaf development in Arabidopsis thaliana: a not-so-gradual process. Dev Cell 22:64–78

    Article  CAS  PubMed  Google Scholar 

  • Balasubramanian R, Karve A, Kandasamy M, Meagher RB, Moore Bd (2007) A role for F-Actin in hexokinase-mediated glucose signaling. Plant Physiol 145:1423–1434

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bolingue W, Rosnoblet C, Leprince O, Vu BL, Aubry C, Buitink J (2010) The MtSNF4b subunit of the sucrose non-fermenting-related kinase complex connects after-ripening and constitutive defense responses in seeds of Medicago truncatula. Plant J 61:792–803

    Article  CAS  PubMed  Google Scholar 

  • Bouly J-P, Gissot L, Lessard P, Kreis M, Thomas M (1999) Arabidopsis thaliana proteins related to the yeast SIP and SNF4 interact with AKINα1, an SNF1-like protein kinase. Plant J 18:541–550

    Article  CAS  PubMed  Google Scholar 

  • Bradford KJ, Downie AB, Gee OH, Alvarado V, Yang H, Dahal P (2003) Abscisic acid and gibberellin differentially regulate expression of genes of the SNF1-related kinase complex in tomato seeds. Plant Physiol 132:1560–1576

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Broeckx T, Hulsmans S, Rolland F (2016) The plant energy sensor: evolutionary conservation and divergence of SnRK1 structure, regulation, and function. J Exp Bot 67:6215–6252

    Article  CAS  PubMed  Google Scholar 

  • Cho Y-H, Yoo S-D, Sheen J (2006) Regulatory functions of nuclear hexokinase1 complex in glucose signaling. Cell 127:579–589

    Article  CAS  PubMed  Google Scholar 

  • Claeyssen É, Rivoal J (2007) Isozymes of plant hexokinase: occurrence, properties and functions. Phytochemistry 68:709–731

    Article  CAS  PubMed  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  CAS  PubMed  Google Scholar 

  • Dai N, Schaffer A, Petreikov M, Shahak Y, Giller Y, Ratner K, Levine A, Granot D (1999) Overexpression of Arabidopsis hexokinase in tomato plants inhibits growth, reduces photosynthesis, and induces rapid senescence. Plant Cell 11:1253–1266

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Damari-Weissler H, Ginzburg A, Gidoni D, Mett A, Krassovskaya I, Weber APM, Belausov E, Granot D (2007) Spinach SoHXK1 is a mitochondria-associated hexokinase. Planta 226:1053–1058

    Article  CAS  PubMed  Google Scholar 

  • Donnelly PM, Bonetta D, Tsukaya H, Dengler RE, Dengler NG (1999) Cell cycling and cell enlargement in developing leaves of Arabidopsis. Dev Biol 215:407–419

    Article  CAS  PubMed  Google Scholar 

  • Eloy NB, Gonzalez N, Van Leene J, Maleux K, Vanhaeren H, De Milde L, Dhondt S, Vercruysse L, Witters E, Mercier R, Cromer L, Beemster GTS, Remaut H, Van Montagu MCE, De Jaeger G, Ferreira PCG, Inzé D (2012) SAMBA, a plant-specific anaphase-promoting complex/cyclosome regulator is involved in early development and A-type cyclin stabilization. Proc Natl Acad Sci USA 109:13853–13858

    Article  PubMed  PubMed Central  Google Scholar 

  • Emanuelle S, Hossain MI, Moller IE, Pedersen HL, van de Meene AML, Doblin MS, Koay A, Oakhill JS, Scott JW, Willats WGT, Kemp BE, Bacic A, Gooley PR, Stapleton DI (2015) SnRK1 from Arabidopsis thaliana is an atypical AMPK. Plant J 82:183–192

    Article  CAS  PubMed  Google Scholar 

  • Fang L, Hou X, Lee LYC, Liu L, Yan X, Yu H (2011) AtPV42a and AtPV42b redundantly regulate reproductive development in Arabidopsis thaliana. PLoS ONE 6:e19033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Feng J, Zhao S, Chen X, Wang W, Dong W, Chen J, Shen J-R, Liu L, Kuang T (2015) Biochemical and structural study of Arabidopsis hexokinase 1. Acta Crystallogr Section D D71:367–375

    Article  CAS  Google Scholar 

  • Fernie AR, Carrari F, Sweetlove LJ (2004) Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport. Curr Opin Plant Biol 7:254–261

    Article  CAS  PubMed  Google Scholar 

  • Fettke J, Fernie AR (2015) Intracellular and cell-to-apoplast compartmentation of carbohydrate metabolism. Trends Plant Sci 20:490–497

    Article  CAS  PubMed  Google Scholar 

  • Geisler M, Nadeau J, Sack FD (2000) Oriented asymmetric divisions that generate the stomatal spacing pattern in Arabidopsis are disrupted by the too many mouths mutation. Plant Cell 12:2075–2086

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gissot L, Polge C, Jossier M, Girin T, Bouly J-P, Kreis M, Thomas M (2006) AKINβγ contributes to SnRK1 heterotrimeric complexes and interacts with two proteins implicated in plant pathogen resistance through its KIS/GBD sequence. Plant Physiol 142:931–944

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gonzalez N, Vanhaeren H, Inzé D (2012) Leaf size control: complex coordination of cell division and expansion. Trends Plant Sci 17:332–340

    Article  CAS  PubMed  Google Scholar 

  • Granot D (2008) Putting plant hexokinases in their proper place. Phytochemistry 69:2649–2654

    Article  CAS  PubMed  Google Scholar 

  • Heazlewood JL, Tonti-Filippini JS, Gout AM, Day DA, Whelan J, Millar AH (2004) Experimental analysis of the Arabidopsis mitochondrial proteome highlights signaling and regulatory components, provides assessment of targeting prediction programs, and indicates plant-specific mitochondrial proteins. Plant Cell 16:241–256

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Heinrichs L, Schmitz J, Flügge U-I, Häusler RE (2012) The mysterious rescue of adg1-1/tpt-2-an Arabidopsis thaliana double mutant impaired in acclimation to high light-by exogenously supplied sugars. Front Plant Sci 3:265

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jang J-C, Léon P, Zhou L, Sheen J (1997) Hexokinase as a sugar sensor in higher plants. Plant Cell 9:5–19

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Karimi M, Bleys A, Vanderhaeghen R, Hilson P (2007a) Building blocks for plant gene assembly. Plant Physiol 145:1183–1191

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Karimi M, Depicker A, Hilson P (2007b) Recombinational cloning with plant Gateway vectors. Plant Physiol 145:1144–1154

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Karve A, Rauh BL, Xia X, Kandasamy M, Meagher RB, Sheen J, Moore Bd (2008) Expression and evolutionary features of the hexokinase gene family in Arabidopsis. Planta 228:411–425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kelly G, David-Schwartz R, Sade N, Moshelion M, Levi A, Alchanatis V, Granot D (2012) The Pitfalls of transgenic selection and new roles of AtHXK1: a high level of AtHXK1 expression uncouples hexokinase1-dependent sugar signaling from exogenous sugar. Plant Physiol 159:47–51

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim Y-M, Heinzel N, Giese J-O, Koeber J, Melzer M, Rutten T, Von Wirén N, Sonnewald U, Hajirezaei M-R (2013) A dual role of tobacco hexokinase 1 in primary metabolism and sugar sensing. Plant Cell Environ 36:1311–1327

    Article  CAS  PubMed  Google Scholar 

  • Lastdrager J, Hanson J, Smeekens S (2014) Sugar signals and the control of plant growth and development. J Exp Bot 65:799–807

    Article  CAS  PubMed  Google Scholar 

  • Millar AH, Whelan J, Soole KL, Day DA (2011) Organization and regulation of mitochondrial respiration in plants. Annu Rev Plant Biol 62:79–104

    Article  CAS  PubMed  Google Scholar 

  • Moore B, Zhou L, Rolland F, Hall Q, Cheng W-H, Liu Y-X, Hwang I, Jones T, Sheen J (2003) Role of the Arabidopsis glucose sensor HXK1 in nutrient, light, and hormonal signaling. Science 300:332–336

    Article  CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Oliva M, Dunand C (2007) Waving and skewing: how gravity and the surface of growth media affect root development in Arabidopsis. New Phytol 176:37–43

    Article  CAS  PubMed  Google Scholar 

  • Ramon M, Ruelens P, Li Y, Sheen J, Geuten K, Rolland F (2013) The hybrid Four-CBS-Domain KINβγ subunit functions as the canonical subunit of the plant energy sensor SnRK1. Plant J 75:11–25

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rasmusson AG, Fernie AR, van Dongen JT (2009) Alternative oxidase: a defence against metabolic fluctuations? Physiol Plant 137:371–382

    Article  CAS  PubMed  Google Scholar 

  • Rolland F, Baena-Gonzalez E, Sheen J (2006) Sugar sensing and signaling in plants: conserved and novel mechanisms. Annu Rev Plant Biol 57:675–709

    Article  CAS  PubMed  Google Scholar 

  • Rosnoblet C, Aubry C, Leprince O, Vu BL, Rogniaux H, Buitink J (2007) The regulatory gamma subunit SNF4b of the sucrose non-fermenting-related kinase complex is involved in longevity and stachyose accumulation during maturation of Medicago truncatula seeds. Plant J 51:47–59

    Article  CAS  PubMed  Google Scholar 

  • Sheen J (2014) Master regulators in plant glucose signaling networks. J Plant Biol 57:67–79

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shimada TL, Shimada T, Hara-Nishimura I (2010) A rapid and non-destructive screenable marker, FAST, for identifying transformed seeds of Arabidopsis thaliana. Plant J 61:519–528

    Article  CAS  PubMed  Google Scholar 

  • Van Leene J, Stals H, Eeckhout D, Persiau G, Van De Slijke E, Van Isterdael G, De Clercq A, Bonnet E, Laukens K, Remmerie N, Henderickx K, De Vijlder T, Abdelkrim A, Pharazyn A, Van Onckelen H, Inzé D, Witters E, De Jaeger G (2007) A tandem affinity purification-based technology platform to study the cell cycle interactome in Arabidopsis thaliana. Mol Cell Proteomics 6:1226–1238

    Article  CAS  PubMed  Google Scholar 

  • Van Leene J, Witters E, Inzé D, De Jaeger G (2008) Boosting tandem affinity purification of plant protein complexes. Trends Plant Sci 13:517–520

    Article  CAS  PubMed  Google Scholar 

  • Van Aken O, Giraud E, Clifton R, Whelan J (2009) Alternative oxidase: a target and regulator of stress responses. Physiol Plant 137:354–361

    Article  CAS  PubMed  Google Scholar 

  • Van Leene J, Eeckhout D, Persiau G, Van De Slijke E, Geerinck J, Van Isterdael G, Witters E, De Jaeger G (2011) Isolation of transcription factor complexes from Arabidopsis cell suspension cultures by tandem affinity purification. Methods Mol Biol 754:195–218

    Article  CAS  PubMed  Google Scholar 

  • Van Leene J, Eeckhout D, Cannoot B, De Winne N, Persiau G, Van De Slijke E, Vercruysse L, Dedecker M, Verkest A, Vandepoele K, Martens L, Witters E, Gevaert K, De Jaeger G (2015) An improved toolbox to unravel the plant cellular machinery by tandem affinity purification of Arabidopsis protein complexes. Nat Protoc 10:169–187

    Article  PubMed  Google Scholar 

  • Van Dingenen J, De Milde L, Vermeersch M, Maleux K, De Rycke RM, De Bruyne M, Storme V, Gonzalez N, Dhondt S, Inzé D (2016) Chloroplasts are central players in sugar-induced leaf growth. Plant Physiol 171:590–605

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vercruyssen L, Verkest A, Gonzalez N, Heyndrickx KS, Eeckhout D, Han S-K, Jégu T, Archacki R, Van Leene J, Andriankaja M, De Bodt S, Abeel T, Coppens F, Dhondt S, De Milde L, Vermeersch M, Maleux K, Gevaert K, Jerzmanowski A, Benhamed M, Wagner D, Vandepoele K, De Jaeger G, Inzé D (2014) ANGUSTIFOLIA3 binds to SWI/SNF chromatin remodeling complexes to regulate transcription during Arabidopsis leaf development. Plant Cell 26:210–229

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xiao B, Sanders MJ, Underwood E, Heath R, Mayer FV, Carmena D, Jing C, Walker PA, Eccleston JF, Haire LF, Saiu P, Howell SA, Aasland R, Martin SR, Carling D, Gamblin SJ (2011) Structure of mammalian AMPK and its regulation by ADP. Nature 472:230–233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xiong Y, Sheen J (2014) The role of target of rapamycin signaling networks in plant growth and metabolism. Plant Physiol 164:499–512

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yoo S-D, Cho Y-H, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2:1565–1572

    Article  CAS  PubMed  Google Scholar 

  • Zhang B, Carrie C, Ivanova A, Narsai R, Murcha MW, Duncan O, Wang Y, Law SR, Albrecht V, Pogson B, Giraud E, Van Aken O, Whelan J (2012) LETM proteins play a role in the accumulation of mitochondrially encoded proteins in Arabidopsis thaliana and AtLETM2 displays parent of origin effects. J Biol Chem 287:41757–41773

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou L, Jang J-c, Jones TL, Sheen J (1998) Glucose and ethylene signal transduction crosstalk revealed by an Arabidopsis glucose-insensitive mutant. Proc Natl Acad Sci USA 95:10294–10299

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank all colleagues of the Systems Biology of Yield research group for many fruitful discussions. Special thanks to Riet De Rycke and Michiel De Bruyne for performing the TEM experiments, Véronique Storme for her help with the statistical analysis, Wim Dejonghe for confocal imaging assistance and Annick Bleys for her help in preparing the manuscript. This work was supported by Ghent University (‘Bijzonder Onderzoeksfonds Methusalem Project’ no. BOF08/01M00408) and the Research Foundation-Flanders (FWO project G046512N “Controle van de bladgroei door suiker-en energiesignalering”).

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Authors and Affiliations

  1. Center for Plant Systems Biology, VIB-Ghent University, Technologiepark 927, 9052, Gent, Belgium

    Judith Van Dingenen, Mattias Vermeersch, Liesbeth De Milde, Nancy De Winne, Jelle Van Leene, Nathalie Gonzalez, Stijn Dhondt, Geert De Jaeger & Dirk Inzé

  2. Department of Plant Biotechnology and Bioinformatics, Ghent University, Gent, Belgium

    Judith Van Dingenen, Mattias Vermeersch, Liesbeth De Milde, Nancy De Winne, Jelle Van Leene, Nathalie Gonzalez, Stijn Dhondt, Geert De Jaeger & Dirk Inzé

  3. Laboratory of Molecular Plant Biology, KU Leuven Department of Biology, Kasteelpark Arenberg 31, 3001, Leuven, Belgium

    Sander Hulsmans & Filip Rolland

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  1. Judith Van Dingenen
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  2. Mattias Vermeersch
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  3. Liesbeth De Milde
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  4. Sander Hulsmans
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  6. Jelle Van Leene
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  7. Nathalie Gonzalez
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  9. Geert De Jaeger
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  10. Filip Rolland
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  11. Dirk Inzé
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Contributions

JVD performed experiments, analyzed data and is the main author; MV and LDM assisted experiments; SH performed the BiFC experiments; JVL, NDW and GDJ performed the TAP experiments; FR, NG, SD and JVL were involved in discussions throughout the project; NG, SD, FR and DI contributed to the writing of the manuscript. GDJ and DI supervised the project.

Corresponding author

Correspondence to Dirk Inzé.

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Van Dingenen, J., Vermeersch, M., De Milde, L. et al. The role of HEXOKINASE1 in Arabidopsis leaf growth. Plant Mol Biol 99, 79–93 (2019). https://doi.org/10.1007/s11103-018-0803-0

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