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Structure, expression, and functional analysis of the hexokinase gene family in rice (Oryza sativa L.)

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Abstract

Hexokinase (HXK) is a dual-function enzyme that both phosphorylates hexose to form hexose 6−phosphate and plays an important role in sugar sensing and signaling. To investigate the roles of hexokinases in rice growth and development, we analyzed rice sequence databases and isolated ten rice hexokinase cDNAs, OsHXK1 (Oryza sativa Hexokinase 1) through OsHXK10. With the exception of the single-exon gene OsHXK1, the OsHXKs all have a highly conserved genomic structure consisting of nine exons and eight introns. Gene expression profiling revealed that OsHXK2 through OsHXK9 are expressed ubiquitously in various organs, whereas OsHXK10 expression is pollen-specific. Sugars induced the expression of three OsHXKs, OsHXK2, OsHXK5, and OsHXK6, in excised leaves, while suppressing OsHXK7 expression in excised leaves and immature seeds. The hexokinase activity of the OsHXKs was confirmed by functional complementation of the hexokinase-deficient yeast strain YSH7.4-3C (hxk1, hxk2, glk1). OsHXK4 was able to complement this mutant only after the chloroplast-transit peptide was removed. The subcellular localization of OsHXK4 and OsHXK7, observed with green fluorescent protein (GFP) fusion constructs, indicated that OsHXK4 is a plastid-stroma-targeted hexokinase while OsHXK7 localizes to the cytosol.

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Abbreviations

cM:

Kosambi values

DAF:

Days after flowering

OsHXK:

Rice hexokinase

UDT1 :

Undeveloped tapetum 1

UTR:

Untranslated region

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Acknowledgements

We thank Dr. Wolf B. Frommer (Carnegie Institution, USA) for the yeast shuttle vector pDR196 and Dr. Yeon-Il Park (Chungnam National University, Korea), Dr. Jong-Min Nam (California Institute of Technology, USA) and Dr. Sangtae Kim (University of Florida, USA) for helpful discussions. We also thank the members of the Plant Metabolism Research Center (PMRC) for helpful discussions. This work was supported, in part, by grants from the SRC program of MOST/KOSEF (R11-2000-081) through the Plant Metabolism Research Center; from the Biogreen 21 Program, Rural Development Administration, from the Crop Functional Genomic Center (CG1422 and CG1111), the 21 Century Frontier Program, and from the BK21 program, Ministry of Education.

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Authors and Affiliations

  1. Plant Metabolism Research Center & Graduate School of Biotechnology, Kyung Hee University, 449-701, Yongin, Republic of South Korea

    Jung-Il Cho, Nayeon Ryoo, Seho Ko, Sang-Kyu Lee, Junok Lee, Youn-Hyung Lee, Seong Hee Bhoo, Tae-Ryong Hahn & Jong-Seong Jeon

  2. Division of Molecular and Life Sciences, Pohang University of Science and Technology, 790-784, Pohang, Republic of South Korea

    Ki-Hong Jung & Gynheung An

  3. Functional Biology, Katholieke Universiteit Leuven, Plantkunde en Microbiologie, Kasteelpark Arenberg 31, 3001, Heverlee, Belgium

    Joris Winderickx

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  1. Jung-Il Cho
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Cho, JI., Ryoo, N., Ko, S. et al. Structure, expression, and functional analysis of the hexokinase gene family in rice (Oryza sativa L.). Planta 224, 598–611 (2006). https://doi.org/10.1007/s00425-006-0251-y

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