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. 2000 Feb 1;345(Pt 3):673–680.

LKB1, a novel serine/threonine protein kinase and potential tumour suppressor, is phosphorylated by cAMP-dependent protein kinase (PKA) and prenylated in vivo.

S P Collins 1, J L Reoma 1, D M Gamm 1, M D Uhler 1
PMCID: PMC1220803  PMID: 10642527

Abstract

Peutz-Jeghers syndrome (PJS) is an autosomal dominant disease characterized by melanocytic macules, hamartomatous polyps and an increased risk for numerous cancers. The human LKB1 (hLKB1) gene encodes a serine/threonine protein kinase that is deficient in the majority of patients with PJS. The murine LKB1 (mLKB1) cDNA was isolated, sequenced and shown to produce a 2.4-kb transcript encoding a 436 amino acid protein with 90% identity with hLKB1. RNA blot and RNase-protection analysis revealed that mLKB1 mRNA is expressed in all tissues and cell lines examined. The widespread expression of LKB1 transcripts is consistent with the elevated risk of multiple cancer types in PJS patients. The predicted LKB1 protein sequence terminates with a conserved prenylation motif (Cys(433)-Lys-Gln-Gln(436)) directly downstream from a consensus cAMP-dependent protein kinase (PKA) phosphorylation site (Arg(428)-Arg-Leu-Ser(431)). The expression of enhanced green fluorescent protein (EGFP)-mLKB1 chimaeras demonstrated that LKB1 possesses a functional prenylation motif that is capable of targeting EGFP to cellular membranes. Mutation of Cys(433) to an alanine residue, but not phosphorylation by PKA, blocked membrane localization. These findings suggest that PKA does phosphorylate LKB1, although this phosphorylation does not alter the cellular localization of LKB1.

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Selected References

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  1. Adamson P., Marshall C. J., Hall A., Tilbrook P. A. Post-translational modifications of p21rho proteins. J Biol Chem. 1992 Oct 5;267(28):20033–20038. [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Avizienyte E., Loukola A., Roth S., Hemminki A., Tarkkanen M., Salovaara R., Arola J., Bützow R., Husgafvel-Pursiainen K., Kokkola A. LKB1 somatic mutations in sporadic tumors. Am J Pathol. 1999 Mar;154(3):677–681. doi: 10.1016/S0002-9440(10)65314-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Avizienyte E., Roth S., Loukola A., Hemminki A., Lothe R. A., Stenwig A. E., Fosså S. D., Salovaara R., Aaltonen L. A. Somatic mutations in LKB1 are rare in sporadic colorectal and testicular tumors. Cancer Res. 1998 May 15;58(10):2087–2090. [PubMed] [Google Scholar]
  5. Ballestero R. P., Wilmot G. R., Leski M. L., Uhler M. D., Agranoff B. W. Isolation of cDNA clones encoding RICH: a protein induced during goldfish optic nerve regeneration with homology to mammalian 2',3'-cyclic-nucleotide 3'-phosphodiesterases. Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8621–8625. doi: 10.1073/pnas.92.19.8621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bignell G. R., Barfoot R., Seal S., Collins N., Warren W., Stratton M. R. Low frequency of somatic mutations in the LKB1/Peutz-Jeghers syndrome gene in sporadic breast cancer. Cancer Res. 1998 Apr 1;58(7):1384–1386. [PubMed] [Google Scholar]
  7. Boardman L. A., Thibodeau S. N., Schaid D. J., Lindor N. M., McDonnell S. K., Burgart L. J., Ahlquist D. A., Podratz K. C., Pittelkow M., Hartmann L. C. Increased risk for cancer in patients with the Peutz-Jeghers syndrome. Ann Intern Med. 1998 Jun 1;128(11):896–899. doi: 10.7326/0003-4819-128-11-199806010-00004. [DOI] [PubMed] [Google Scholar]
  8. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  10. Collins S. P., Uhler M. D. Characterization of PKIgamma, a novel isoform of the protein kinase inhibitor of cAMP-dependent protein kinase. J Biol Chem. 1997 Jul 18;272(29):18169–18178. doi: 10.1074/jbc.272.29.18169. [DOI] [PubMed] [Google Scholar]
  11. Dell'Acqua M. L., Scott J. D. Protein kinase A anchoring. J Biol Chem. 1997 May 16;272(20):12881–12884. doi: 10.1074/jbc.272.20.12881. [DOI] [PubMed] [Google Scholar]
  12. Dunphy J. T., Linder M. E. Signalling functions of protein palmitoylation. Biochim Biophys Acta. 1998 Dec 8;1436(1-2):245–261. doi: 10.1016/s0005-2760(98)00130-1. [DOI] [PubMed] [Google Scholar]
  13. Fearon E. R. Human cancer syndromes: clues to the origin and nature of cancer. Science. 1997 Nov 7;278(5340):1043–1050. doi: 10.1126/science.278.5340.1043. [DOI] [PubMed] [Google Scholar]
  14. Giardiello F. M., Welsh S. B., Hamilton S. R., Offerhaus G. J., Gittelsohn A. M., Booker S. V., Krush A. J., Yardley J. H., Luk G. D. Increased risk of cancer in the Peutz-Jeghers syndrome. N Engl J Med. 1987 Jun 11;316(24):1511–1514. doi: 10.1056/NEJM198706113162404. [DOI] [PubMed] [Google Scholar]
  15. Hancock J. F., Magee A. I., Childs J. E., Marshall C. J. All ras proteins are polyisoprenylated but only some are palmitoylated. Cell. 1989 Jun 30;57(7):1167–1177. doi: 10.1016/0092-8674(89)90054-8. [DOI] [PubMed] [Google Scholar]
  16. Hancock J. F., Paterson H., Marshall C. J. A polybasic domain or palmitoylation is required in addition to the CAAX motif to localize p21ras to the plasma membrane. Cell. 1990 Oct 5;63(1):133–139. doi: 10.1016/0092-8674(90)90294-o. [DOI] [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. Hanks S. K., Quinn A. M. Protein kinase catalytic domain sequence database: identification of conserved features of primary structure and classification of family members. Methods Enzymol. 1991;200:38–62. doi: 10.1016/0076-6879(91)00126-h. [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. Hawley S. A., Davison M., Woods A., Davies S. P., Beri R. K., Carling D., Hardie D. G. Characterization of the AMP-activated protein kinase kinase from rat liver and identification of threonine 172 as the major site at which it phosphorylates AMP-activated protein kinase. J Biol Chem. 1996 Nov 1;271(44):27879–27887. doi: 10.1074/jbc.271.44.27879. [DOI] [PubMed] [Google Scholar]
  21. Hemminki A., Markie D., Tomlinson I., Avizienyte E., Roth S., Loukola A., Bignell G., Warren W., Aminoff M., Höglund P. A serine/threonine kinase gene defective in Peutz-Jeghers syndrome. Nature. 1998 Jan 8;391(6663):184–187. doi: 10.1038/34432. [DOI] [PubMed] [Google Scholar]
  22. Hemminki A., Tomlinson I., Markie D., Järvinen H., Sistonen P., Björkqvist A. M., Knuutila S., Salovaara R., Bodmer W., Shibata D. Localization of a susceptibility locus for Peutz-Jeghers syndrome to 19p using comparative genomic hybridization and targeted linkage analysis. Nat Genet. 1997 Jan;15(1):87–90. doi: 10.1038/ng0197-87. [DOI] [PubMed] [Google Scholar]
  23. Huggenvik J. I., Collard M. W., Stofko R. E., Seasholtz A. F., Uhler M. D. Regulation of the human enkephalin promoter by two isoforms of the catalytic subunit of cyclic adenosine 3',5'-monophosphate-dependent protein kinase. Mol Endocrinol. 1991 Jul;5(7):921–930. doi: 10.1210/mend-5-7-921. [DOI] [PubMed] [Google Scholar]
  24. Inglese J., Koch W. J., Caron M. G., Lefkowitz R. J. Isoprenylation in regulation of signal transduction by G-protein-coupled receptor kinases. Nature. 1992 Sep 10;359(6391):147–150. doi: 10.1038/359147a0. [DOI] [PubMed] [Google Scholar]
  25. Jenne D. E., Reimann H., Nezu J., Friedel W., Loff S., Jeschke R., Müller O., Back W., Zimmer M. Peutz-Jeghers syndrome is caused by mutations in a novel serine threonine kinase. Nat Genet. 1998 Jan;18(1):38–43. doi: 10.1038/ng0198-38. [DOI] [PubMed] [Google Scholar]
  26. Johnson L. N., Noble M. E., Owen D. J. Active and inactive protein kinases: structural basis for regulation. Cell. 1996 Apr 19;85(2):149–158. doi: 10.1016/s0092-8674(00)81092-2. [DOI] [PubMed] [Google Scholar]
  27. Khosravi-Far R., Cox A. D., Kato K., Der C. J. Protein prenylation: key to ras function and cancer intervention? Cell Growth Differ. 1992 Jul;3(7):461–469. [PubMed] [Google Scholar]
  28. Kim C. M., Goldstein J. L., Brown M. S. cDNA cloning of MEV, a mutant protein that facilitates cellular uptake of mevalonate, and identification of the point mutation responsible for its gain of function. J Biol Chem. 1992 Nov 15;267(32):23113–23121. [PubMed] [Google Scholar]
  29. Kita T., Brown M. S., Goldstein J. L. Feedback regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in livers of mice treated with mevinolin, a competitive inhibitor of the reductase. J Clin Invest. 1980 Nov;66(5):1094–1100. doi: 10.1172/JCI109938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lennon G., Auffray C., Polymeropoulos M., Soares M. B. The I.M.A.G.E. Consortium: an integrated molecular analysis of genomes and their expression. Genomics. 1996 Apr 1;33(1):151–152. doi: 10.1006/geno.1996.0177. [DOI] [PubMed] [Google Scholar]
  31. Maller J. L., Krebs E. G. Progesterone-stimulated meiotic cell division in Xenopus oocytes. Induction by regulatory subunit and inhibition by catalytic subunit of adenosine 3':5'-monophosphate-dependent protein kinase. J Biol Chem. 1977 Mar 10;252(5):1712–1718. [PubMed] [Google Scholar]
  32. Mehenni H., Gehrig C., Nezu J., Oku A., Shimane M., Rossier C., Guex N., Blouin J. L., Scott H. S., Antonarakis S. E. Loss of LKB1 kinase activity in Peutz-Jeghers syndrome, and evidence for allelic and locus heterogeneity. Am J Hum Genet. 1998 Dec;63(6):1641–1650. doi: 10.1086/302159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Muramatsu M., Kaibuchi K., Arai K. A protein kinase C cDNA without the regulatory domain is active after transfection in vivo in the absence of phorbol ester. Mol Cell Biol. 1989 Feb;9(2):831–836. doi: 10.1128/mcb.9.2.831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Otto J. C., Casey P. J. The hepatitis delta virus large antigen is farnesylated both in vitro and in animal cells. J Biol Chem. 1996 Mar 1;271(9):4569–4572. doi: 10.1074/jbc.271.9.4569. [DOI] [PubMed] [Google Scholar]
  35. Pitcher J. A., Freedman N. J., Lefkowitz R. J. G protein-coupled receptor kinases. Annu Rev Biochem. 1998;67:653–692. doi: 10.1146/annurev.biochem.67.1.653. [DOI] [PubMed] [Google Scholar]
  36. Reiss Y., Stradley S. J., Gierasch L. M., Brown M. S., Goldstein J. L. Sequence requirement for peptide recognition by rat brain p21ras protein farnesyltransferase. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):732–736. doi: 10.1073/pnas.88.3.732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Su J. Y., Erikson E., Maller J. L. Cloning and characterization of a novel serine/threonine protein kinase expressed in early Xenopus embryos. J Biol Chem. 1996 Jun 14;271(24):14430–14437. doi: 10.1074/jbc.271.24.14430. [DOI] [PubMed] [Google Scholar]
  38. Uhler M. D. Cloning and expression of a novel cyclic GMP-dependent protein kinase from mouse brain. J Biol Chem. 1993 Jun 25;268(18):13586–13591. [PubMed] [Google Scholar]
  39. Wang Z. J., Taylor F., Churchman M., Norbury G., Tomlinson I. Genetic pathways of colorectal carcinogenesis rarely involve the PTEN and LKB1 genes outside the inherited hamartoma syndromes. Am J Pathol. 1998 Aug;153(2):363–366. doi: 10.1016/S0002-9440(10)65579-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Willumsen B. M., Cox A. D., Solski P. A., Der C. J., Buss J. E. Novel determinants of H-Ras plasma membrane localization and transformation. Oncogene. 1996 Nov 7;13(9):1901–1909. [PubMed] [Google Scholar]
  41. Ylikorkala A., Avizienyte E., Tomlinson I. P., Tiainen M., Roth S., Loukola A., Hemminki A., Johansson M., Sistonen P., Markie D. Mutations and impaired function of LKB1 in familial and non-familial Peutz-Jeghers syndrome and a sporadic testicular cancer. Hum Mol Genet. 1999 Jan;8(1):45–51. doi: 10.1093/hmg/8.1.45. [DOI] [PubMed] [Google Scholar]
  42. Zhang F. L., Casey P. J. Protein prenylation: molecular mechanisms and functional consequences. Annu Rev Biochem. 1996;65:241–269. doi: 10.1146/annurev.bi.65.070196.001325. [DOI] [PubMed] [Google Scholar]

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