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J. Biol. Chem., Vol. 282, Issue 14, 10472-10479, April 6, 2007

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DNA Sequences from Arabidopsis, Which Encode Protein Kinases and Function as Upstream Regulators of Snf1 in Yeast^*

Sandra Hey, Hubert Mayerhofer¹, Nigel G. Halford, and J. Richard Dickinson²

From the Department of Crop Performance and Improvement, Rothamsted Research, Harpenden AL5 2JQ, United Kingdom and the Cardiff School of Biosciences, Cardiff University, P. O. Box 915, Cardiff CF10 3TL, United Kingdom

Sucrose nonfermenting-1 (Snf1)-related protein kinase-1 (SnRK1) of plants is a global regulator of carbon metabolism through the modulation of enzyme activity and gene expression. It is structurally and functionally related to the yeast protein kinase, Snf1, and to mammalian AMP-activated protein kinase. Two DNA sequences from Arabidopsis thaliana, previously known only by their data base accession numbers of NM_ 125448.3 (protein ID NP_200863 [GenBank] ) and NM_114393 [GenBank] .3 (protein ID NP_566876 [GenBank] ) each functionally complemented a Saccharomyces cerevisiae elm1 sak1 tos3 triple mutant. This indicates that the Arabidopsis proteins are able to substitute for one of the missing yeast upstream kinases, which are required for activity of Snf1. Both plant proteins were shown to phosphorylate a peptide with the amino acid sequence of the phosphorylation site in the T-loop of SnRK1 and by inference SnRK1 in Arabidopsis. The proteins encoded by NM_125448 [GenBank] .3 and NM_114393 [GenBank] .3 have been named AtSnAK1 and AtSnAK2 (Arabidopsis thaliana SnRK1-activating kinase), respectively. We believe this is the first time that upstream activators of SnRK1 have been described in any plant species.

Received for publication, December 7, 2006 , and in revised form, January 19, 2007.

The nucleotide sequence(s) reported in this paper has been submitted to GenBank^TM/EBI Data Bank with the accession number(s) AM489730 [GenBank] and AM489731 [GenBank] .

^* This work was supported by a grant from the Biotechnology and Biological Sciences Research Council (BBSRC) of the United Kingdom (to Rothamsted Research). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Recipient of a studentship under the European Union's ERASMUS program.

² To whom correspondence should be addressed. Tel.: 44-2920-875762; Fax: 44-2920-874305; E-mail: Dickinson{at}cardiff.ac.uk.

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