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. 2001 Apr 15;355(Pt 2):297–305. doi: 10.1042/0264-6021:3550297

Identification and characterization of a novel sucrose-non-fermenting protein kinase/AMP-activated protein kinase-related protein kinase, SNARK.

D L Lefebvre 1, Y Bai 1, N Shahmolky 1, M Sharma 1, R Poon 1, D J Drucker 1, C F Rosen 1
PMCID: PMC1221739  PMID: 11284715

Abstract

Subtraction hybridization after the exposure of keratinocytes to ultraviolet radiation identified a differentially expressed cDNA that encodes a protein of 630 amino acid residues possessing significant similarity to the catalytic domain of the sucrose-non-fermenting protein kinase (SNF1)/AMP-activated protein kinase (AMPK) family of serine/threonine protein kinases. Northern blotting and reverse-transcriptase-mediated PCR demonstrated that mRNA transcripts for the SNF1/AMPK-related kinase (SNARK) were widely expressed in rodent tissues. The SNARK gene was localized to human chromosome 1q32 by fluorescent in situ hybridization. SNARK was translated in vitro to yield a single protein band of approx. 76 kDa; Western analysis of transfected baby hamster kidney (BHK) cells detected two SNARK-immunoreactive bands of approx. 76-80 kDa. SNARK was capable of autophosphorylation in vitro; immunoprecipitated SNARK exhibited phosphotransferase activity with the synthetic peptide substrate HMRSAMSGLHLVKRR (SAMS) as a kinase substrate. SNARK activity was significantly increased by AMP and 5-amino-4-imidazolecarboxamide riboside (AICAriboside) in rat keratinocyte cells, implying that SNARK might be activated by an AMPK kinase-dependent pathway. Furthermore, glucose deprivation increased SNARK activity 3-fold in BHK fibroblasts. These findings identify SNARK as a glucose- and AICAriboside-regulated member of the AMPK-related gene family that represents a new candidate mediator of the cellular response to metabolic stress.

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Selected References

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  1. Alderson A., Sabelli P. A., Dickinson J. R., Cole D., Richardson M., Kreis M., Shewry P. R., Halford N. G. Complementation of snf1, a mutation affecting global regulation of carbon metabolism in yeast, by a plant protein kinase cDNA. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8602–8605. doi: 10.1073/pnas.88.19.8602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker W., Heukelbach J., Kentrup H., Joost H. G. Molecular cloning and characterization of a novel mammalian protein kinase harboring a homology domain that defines a subfamily of serine/threonine kinases. Eur J Biochem. 1996 Feb 1;235(3):736–743. doi: 10.1111/j.1432-1033.1996.00736.x. [DOI] [PubMed] [Google Scholar]
  3. Bouly J. P., Gissot L., Lessard P., Kreis M., Thomas M. Arabidopsis thaliana proteins related to the yeast SIP and SNF4 interact with AKINalpha1, an SNF1-like protein kinase. Plant J. 1999 Jun;18(5):541–550. doi: 10.1046/j.1365-313x.1999.00476.x. [DOI] [PubMed] [Google Scholar]
  4. Bracchi V., Langsley G., Thélu J., Eling W., Ambroise-Thomas P. PfKIN, an SNF1 type protein kinase of Plasmodium falciparum predominantly expressed in gametocytes. Mol Biochem Parasitol. 1996 Feb-Mar;76(1-2):299–303. doi: 10.1016/0166-6851(96)02564-9. [DOI] [PubMed] [Google Scholar]
  5. Carling D., Aguan K., Woods A., Verhoeven A. J., Beri R. K., Brennan C. H., Sidebottom C., Davison M. D., Scott J. Mammalian AMP-activated protein kinase is homologous to yeast and plant protein kinases involved in the regulation of carbon metabolism. J Biol Chem. 1994 Apr 15;269(15):11442–11448. [PubMed] [Google Scholar]
  6. Carlson M. Glucose repression in yeast. Curr Opin Microbiol. 1999 Apr;2(2):202–207. doi: 10.1016/S1369-5274(99)80035-6. [DOI] [PubMed] [Google Scholar]
  7. Celenza J. L., Carlson M. A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science. 1986 Sep 12;233(4769):1175–1180. doi: 10.1126/science.3526554. [DOI] [PubMed] [Google Scholar]
  8. Chen C. A., Okayama H. Calcium phosphate-mediated gene transfer: a highly efficient transfection system for stably transforming cells with plasmid DNA. Biotechniques. 1988 Jul-Aug;6(7):632–638. [PubMed] [Google Scholar]
  9. Corton J. M., Gillespie J. G., Hawley S. A., Hardie D. G. 5-aminoimidazole-4-carboxamide ribonucleoside. A specific method for activating AMP-activated protein kinase in intact cells? Eur J Biochem. 1995 Apr 15;229(2):558–565. doi: 10.1111/j.1432-1033.1995.tb20498.x. [DOI] [PubMed] [Google Scholar]
  10. Dale S., Wilson W. A., Edelman A. M., Hardie D. G. Similar substrate recognition motifs for mammalian AMP-activated protein kinase, higher plant HMG-CoA reductase kinase-A, yeast SNF1, and mammalian calmodulin-dependent protein kinase I. FEBS Lett. 1995 Mar 20;361(2-3):191–195. doi: 10.1016/0014-5793(95)00172-6. [DOI] [PubMed] [Google Scholar]
  11. Davies J. P., Yildiz F. H., Grossman A. R. Sac3, an Snf1-like serine/threonine kinase that positively and negatively regulates the responses of Chlamydomonas to sulfur limitation. Plant Cell. 1999 Jun;11(6):1179–1190. doi: 10.1105/tpc.11.6.1179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Davies S. P., Carling D., Hardie D. G. Tissue distribution of the AMP-activated protein kinase, and lack of activation by cyclic-AMP-dependent protein kinase, studied using a specific and sensitive peptide assay. Eur J Biochem. 1989 Dec 8;186(1-2):123–128. doi: 10.1111/j.1432-1033.1989.tb15185.x. [DOI] [PubMed] [Google Scholar]
  13. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  14. Gao G., Widmer J., Stapleton D., Teh T., Cox T., Kemp B. E., Witters L. A. Catalytic subunits of the porcine and rat 5'-AMP-activated protein kinase are members of the SNF1 protein kinase family. Biochim Biophys Acta. 1995 Apr 6;1266(1):73–82. doi: 10.1016/0167-4889(94)00222-z. [DOI] [PubMed] [Google Scholar]
  15. Gardner H. P., Wertheim G. B., Ha S. I., Copeland N. G., Gilbert D. J., Jenkins N. A., Marquis S. T., Chodosh L. A. Cloning and characterization of Hunk, a novel mammalian SNF1-related protein kinase. Genomics. 2000 Jan 1;63(1):46–59. doi: 10.1006/geno.1999.6078. [DOI] [PubMed] [Google Scholar]
  16. Halford N. G., Vicente-Carbajosa J., Sabelli P. A., Shewry P. R., Hannappel U., Kreis M. Molecular analyses of a barley multigene family homologous to the yeast protein kinase gene SNF1. Plant J. 1992 Sep;2(5):791–797. [PubMed] [Google Scholar]
  17. Hanks S. K., Hunter T. Protein kinases 6. The eukaryotic protein kinase superfamily: kinase (catalytic) domain structure and classification. FASEB J. 1995 May;9(8):576–596. [PubMed] [Google Scholar]
  18. Hannappel U., Vicente-Carbajosa J., Barker J. H., Shewry P. R., Halford N. G. Differential expression of two barley SNF1-related protein kinase genes. Plant Mol Biol. 1995 Mar;27(6):1235–1240. doi: 10.1007/BF00020898. [DOI] [PubMed] [Google Scholar]
  19. Hardie D. G., Carling D., Carlson M. The AMP-activated/SNF1 protein kinase subfamily: metabolic sensors of the eukaryotic cell? Annu Rev Biochem. 1998;67:821–855. doi: 10.1146/annurev.biochem.67.1.821. [DOI] [PubMed] [Google Scholar]
  20. Henin N., Vincent M. F., Van den Berghe G. Stimulation of rat liver AMP-activated protein kinase by AMP analogues. Biochim Biophys Acta. 1996 Jun 4;1290(2):197–203. doi: 10.1016/0304-4165(96)00021-9. [DOI] [PubMed] [Google Scholar]
  21. Heyer B. S., Warsowe J., Solter D., Knowles B. B., Ackerman S. L. New member of the Snf1/AMPK kinase family, Melk, is expressed in the mouse egg and preimplantation embryo. Mol Reprod Dev. 1997 Jun;47(2):148–156. doi: 10.1002/(SICI)1098-2795(199706)47:2<148::AID-MRD4>3.0.CO;2-M. [DOI] [PubMed] [Google Scholar]
  22. Higgins D. G., Sharp P. M. Fast and sensitive multiple sequence alignments on a microcomputer. Comput Appl Biosci. 1989 Apr;5(2):151–153. doi: 10.1093/bioinformatics/5.2.151. [DOI] [PubMed] [Google Scholar]
  23. Inglis J. D., Lee M., Hill R. E. Emk, a protein kinase with homologs in yeast maps to mouse chromosome 19. Mamm Genome. 1993;4(7):401–403. doi: 10.1007/BF00360595. [DOI] [PubMed] [Google Scholar]
  24. Ip Y. T., Davis R. J. Signal transduction by the c-Jun N-terminal kinase (JNK)--from inflammation to development. Curr Opin Cell Biol. 1998 Apr;10(2):205–219. doi: 10.1016/s0955-0674(98)80143-9. [DOI] [PubMed] [Google Scholar]
  25. Le Guen L., Thomas M., Bianchi M., Halford N. G., Kreis M. Structure and expression of a gene from Arabidopsis thaliana encoding a protein related to SNF1 protein kinase. Gene. 1992 Oct 21;120(2):249–254. doi: 10.1016/0378-1119(92)90100-4. [DOI] [PubMed] [Google Scholar]
  26. Lichter P., Tang C. J., Call K., Hermanson G., Evans G. A., Housman D., Ward D. C. High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. Science. 1990 Jan 5;247(4938):64–69. doi: 10.1126/science.2294592. [DOI] [PubMed] [Google Scholar]
  27. Muranaka T., Banno H., Machida Y. Characterization of tobacco protein kinase NPK5, a homolog of Saccharomyces cerevisiae SNF1 that constitutively activates expression of the glucose-repressible SUC2 gene for a secreted invertase of S. cerevisiae. Mol Cell Biol. 1994 May;14(5):2958–2965. doi: 10.1128/mcb.14.5.2958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rosen C. F., Gajic D., Drucker D. J. Ultraviolet radiation induction of ornithine decarboxylase in rat keratinocytes. Cancer Res. 1990 May 1;50(9):2631–2635. [PubMed] [Google Scholar]
  29. Rosen C. F., Poon R., Drucker D. J. UVB radiation-activated genes induced by transcriptional and posttranscriptional mechanisms in rat keratinocytes. Am J Physiol. 1995 Apr;268(4 Pt 1):C846–C855. doi: 10.1152/ajpcell.1995.268.4.C846. [DOI] [PubMed] [Google Scholar]
  30. Ruiz J. C., Conlon F. L., Robertson E. J. Identification of novel protein kinases expressed in the myocardium of the developing mouse heart. Mech Dev. 1994 Dec;48(3):153–164. doi: 10.1016/0925-4773(94)90056-6. [DOI] [PubMed] [Google Scholar]
  31. Salt I. P., Johnson G., Ashcroft S. J., Hardie D. G. AMP-activated protein kinase is activated by low glucose in cell lines derived from pancreatic beta cells, and may regulate insulin release. Biochem J. 1998 Nov 1;335(Pt 3):533–539. doi: 10.1042/bj3350533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Salt I., Celler J. W., Hawley S. A., Prescott A., Woods A., Carling D., Hardie D. G. AMP-activated protein kinase: greater AMP dependence, and preferential nuclear localization, of complexes containing the alpha2 isoform. Biochem J. 1998 Aug 15;334(Pt 1):177–187. doi: 10.1042/bj3340177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shahmolky N., Lefebvre D. L., Poon R., Bai Y., Sharma M., Rosen C. F. UVB and gamma-radiation induce the expression of mRNAs encoding the ribosomal subunit L13A in rat keratinocytes. Photochem Photobiol. 1999 Sep;70(3):341–347. [PubMed] [Google Scholar]
  34. Sugden C., Donaghy P. G., Halford N. G., Hardie D. G. Two SNF1-related protein kinases from spinach leaf phosphorylate and inactivate 3-hydroxy-3-methylglutaryl-coenzyme A reductase, nitrate reductase, and sucrose phosphate synthase in vitro. Plant Physiol. 1999 May;120(1):257–274. doi: 10.1104/pp.120.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Takano M., Kajiya-Kanegae H., Funatsuki H., Kikuchi S. Rice has two distinct classes of protein kinase genes related to SNF1 of Saccharomyces cerevisiae, which are differently regulated in early seed development. Mol Gen Genet. 1998 Nov;260(4):388–394. doi: 10.1007/s004380050908. [DOI] [PubMed] [Google Scholar]
  36. Vincent M. F., Marangos P. J., Gruber H. E., Van den Berghe G. Inhibition by AICA riboside of gluconeogenesis in isolated rat hepatocytes. Diabetes. 1991 Oct;40(10):1259–1266. doi: 10.2337/diab.40.10.1259. [DOI] [PubMed] [Google Scholar]
  37. Wang Z., Takemori H., Halder S. K., Nonaka Y., Okamoto M. Cloning of a novel kinase (SIK) of the SNF1/AMPK family from high salt diet-treated rat adrenal. FEBS Lett. 1999 Jun 18;453(1-2):135–139. doi: 10.1016/s0014-5793(99)00708-5. [DOI] [PubMed] [Google Scholar]
  38. Wilson W. A., Hawley S. A., Hardie D. G. Glucose repression/derepression in budding yeast: SNF1 protein kinase is activated by phosphorylation under derepressing conditions, and this correlates with a high AMP:ATP ratio. Curr Biol. 1996 Nov 1;6(11):1426–1434. doi: 10.1016/s0960-9822(96)00747-6. [DOI] [PubMed] [Google Scholar]
  39. Winder W. W., Hardie D. G. Inactivation of acetyl-CoA carboxylase and activation of AMP-activated protein kinase in muscle during exercise. Am J Physiol. 1996 Feb;270(2 Pt 1):E299–E304. doi: 10.1152/ajpendo.1996.270.2.E299. [DOI] [PubMed] [Google Scholar]
  40. Woods A., Munday M. R., Scott J., Yang X., Carlson M., Carling D. Yeast SNF1 is functionally related to mammalian AMP-activated protein kinase and regulates acetyl-CoA carboxylase in vivo. J Biol Chem. 1994 Jul 29;269(30):19509–19515. [PubMed] [Google Scholar]
  41. Young M. E., Radda G. K., Leighton B. Activation of glycogen phosphorylase and glycogenolysis in rat skeletal muscle by AICAR--an activator of AMP-activated protein kinase. FEBS Lett. 1996 Mar 11;382(1-2):43–47. doi: 10.1016/0014-5793(96)00129-9. [DOI] [PubMed] [Google Scholar]
  42. Yusta B., Somwar R., Wang F., Munroe D., Grinstein S., Klip A., Drucker D. J. Identification of glucagon-like peptide-2 (GLP-2)-activated signaling pathways in baby hamster kidney fibroblasts expressing the rat GLP-2 receptor. J Biol Chem. 1999 Oct 22;274(43):30459–30467. doi: 10.1074/jbc.274.43.30459. [DOI] [PubMed] [Google Scholar]
  43. da Silva Xavier G., Leclerc I., Salt I. P., Doiron B., Hardie D. G., Kahn A., Rutter G. A. Role of AMP-activated protein kinase in the regulation by glucose of islet beta cell gene expression. Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4023–4028. doi: 10.1073/pnas.97.8.4023. [DOI] [PMC free article] [PubMed] [Google Scholar]

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