Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1992 Feb;11(2):507–517. doi: 10.1002/j.1460-2075.1992.tb05081.x

Expression of a constitutive form of calcium/calmodulin dependent protein kinase II leads to arrest of the cell cycle in G2.

M D Planas-Silva 1, A R Means 1
PMCID: PMC556481  PMID: 1371461

Abstract

Calcium/calmodulin dependent protein kinase II (CaMKII) is a multifunctional serine/threonine protein kinase. We have created a calcium/calmodulin independent form of this enzyme by truncation. Expression of this enzyme fragment in a rabbit reticulocyte lysate yields a constitutive enzyme with specific activity similar to the activated native enzyme. We have established mammalian cell lines that transiently express this constitutive enzyme using the glucocorticoid-inducible mouse mammary tumor virus long terminal repeat. The transient increase in kinase activity results in a complete cessation of cell cycle progression. This block develops as a consequence of a specific arrest of the cell cycle in G2. During the block, increases in histone H1 kinase activity present in p13 beads or anti-cdc2 immunoprecipitates are seen in parallel with the accumulation of cells at G2, arguing that the arrest is not due to a failure to activate cdc2 as a histone H1 kinase. These results suggest that other changes in serine/threonine protein phosphorylation besides those involved in activation of cdc2 as a histone H1 kinase may be necessary for proper G2-M transition.

Full text

PDF
507

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arion D., Meijer L., Brizuela L., Beach D. cdc2 is a component of the M phase-specific histone H1 kinase: evidence for identity with MPF. Cell. 1988 Oct 21;55(2):371–378. doi: 10.1016/0092-8674(88)90060-8. [DOI] [PubMed] [Google Scholar]
  2. Baitinger C., Alderton J., Poenie M., Schulman H., Steinhardt R. A. Multifunctional Ca2+/calmodulin-dependent protein kinase is necessary for nuclear envelope breakdown. J Cell Biol. 1990 Nov;111(5 Pt 1):1763–1773. doi: 10.1083/jcb.111.5.1763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bartelt D. C., Fidel S., Farber L. H., Wolff D. J., Hammell R. L. Calmodulin-dependent multifunctional protein kinase in Aspergillus nidulans. Proc Natl Acad Sci U S A. 1988 May;85(10):3279–3283. doi: 10.1073/pnas.85.10.3279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ben-David Y., Letwin K., Tannock L., Bernstein A., Pawson T. A mammalian protein kinase with potential for serine/threonine and tyrosine phosphorylation is related to cell cycle regulators. EMBO J. 1991 Feb;10(2):317–325. doi: 10.1002/j.1460-2075.1991.tb07952.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brinkley B. R., Fistel S. H., Marcum J. M., Pardue R. L. Microtubules in cultured cells; indirect immunofluorescent staining with tubulin antibody. Int Rev Cytol. 1980;63:59–95. doi: 10.1016/s0074-7696(08)61757-x. [DOI] [PubMed] [Google Scholar]
  6. Brown N. A., Stofko R. E., Uhler M. D. Induction of alkaline phosphatase in mouse L cells by overexpression of the catalytic subunit of cAMP-dependent protein kinase. J Biol Chem. 1990 Aug 5;265(22):13181–13189. [PubMed] [Google Scholar]
  7. Chafouleas J. G., Bolton W. E., Hidaka H., Boyd A. E., 3rd, Means A. R. Calmodulin and the cell cycle: involvement in regulation of cell-cycle progression. Cell. 1982 Jan;28(1):41–50. doi: 10.1016/0092-8674(82)90373-7. [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. Colbran R. J., Schworer C. M., Hashimoto Y., Fong Y. L., Rich D. P., Smith M. K., Soderling T. R. Calcium/calmodulin-dependent protein kinase II. Biochem J. 1989 Mar 1;258(2):313–325. doi: 10.1042/bj2580313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Connelly P. A., Sisk R. B., Schulman H., Garrison J. C. Evidence for the activation of the multifunctional Ca2+/calmodulin-dependent protein kinase in response to hormones that increase intracellular Ca2+. J Biol Chem. 1987 Jul 25;262(21):10154–10163. [PubMed] [Google Scholar]
  11. Cyert M. S., Thorner J. Putting it on and taking it off: phosphoprotein phosphatase involvement in cell cycle regulation. Cell. 1989 Jun 16;57(6):891–893. doi: 10.1016/0092-8674(89)90325-5. [DOI] [PubMed] [Google Scholar]
  12. Davis F. M., Tsao T. Y., Fowler S. K., Rao P. N. Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2926–2930. doi: 10.1073/pnas.80.10.2926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Draetta G., Beach D. Activation of cdc2 protein kinase during mitosis in human cells: cell cycle-dependent phosphorylation and subunit rearrangement. Cell. 1988 Jul 1;54(1):17–26. doi: 10.1016/0092-8674(88)90175-4. [DOI] [PubMed] [Google Scholar]
  14. Draetta G., Luca F., Westendorf J., Brizuela L., Ruderman J., Beach D. Cdc2 protein kinase is complexed with both cyclin A and B: evidence for proteolytic inactivation of MPF. Cell. 1989 Mar 10;56(5):829–838. doi: 10.1016/0092-8674(89)90687-9. [DOI] [PubMed] [Google Scholar]
  15. Featherstone C., Russell P. Fission yeast p107wee1 mitotic inhibitor is a tyrosine/serine kinase. Nature. 1991 Feb 28;349(6312):808–811. doi: 10.1038/349808a0. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Fong Y. L., Taylor W. L., Means A. R., Soderling T. R. Studies of the regulatory mechanism of Ca2+/calmodulin-dependent protein kinase II. Mutation of threonine 286 to alanine and aspartate. J Biol Chem. 1989 Oct 5;264(28):16759–16763. [PubMed] [Google Scholar]
  18. Freeman R. S., Donoghue D. J. Protein kinases and protooncogenes: biochemical regulators of the eukaryotic cell cycle. Biochemistry. 1991 Mar 5;30(9):2293–2302. doi: 10.1021/bi00223a001. [DOI] [PubMed] [Google Scholar]
  19. Fukunaga K., Goto S., Miyamoto E. Immunohistochemical localization of Ca2+/calmodulin-dependent protein kinase II in rat brain and various tissues. J Neurochem. 1988 Oct;51(4):1070–1078. doi: 10.1111/j.1471-4159.1988.tb03070.x. [DOI] [PubMed] [Google Scholar]
  20. Gautier J., Maller J. L. Cyclin B in Xenopus oocytes: implications for the mechanism of pre-MPF activation. EMBO J. 1991 Jan;10(1):177–182. doi: 10.1002/j.1460-2075.1991.tb07934.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gautier J., Matsukawa T., Nurse P., Maller J. Dephosphorylation and activation of Xenopus p34cdc2 protein kinase during the cell cycle. Nature. 1989 Jun 22;339(6226):626–629. doi: 10.1038/339626a0. [DOI] [PubMed] [Google Scholar]
  22. Gould K. L., Nurse P. Tyrosine phosphorylation of the fission yeast cdc2+ protein kinase regulates entry into mitosis. Nature. 1989 Nov 2;342(6245):39–45. doi: 10.1038/342039a0. [DOI] [PubMed] [Google Scholar]
  23. Hamilton B. J., DeFranco D. Glucocorticoid and cAMP induction mechanisms are differentially affected by the p85gag-mos oncoprotein. Proc Natl Acad Sci U S A. 1989 Jan;86(2):597–601. doi: 10.1073/pnas.86.2.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hanley R. M., Payne M. E., Cruzalegui F., Christenson M. A., Means A. R. Sequence of the cDNA for the alpha subunit of calmodulin kinase II from mouse brain. Nucleic Acids Res. 1989 May 25;17(10):3992–3992. doi: 10.1093/nar/17.10.3992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jaggi R., Salmons B., Muellener D., Groner B. The v-mos and H-ras oncogene expression represses glucocorticoid hormone-dependent transcription from the mouse mammary tumor virus LTR. EMBO J. 1986 Oct;5(10):2609–2616. doi: 10.1002/j.1460-2075.1986.tb04541.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jefferson A. B., Schulman H. Phosphorylation of microtubule-associated protein-2 in GH3 cells. Regulation by cAMP and by calcium. J Biol Chem. 1991 Jan 5;266(1):346–354. [PubMed] [Google Scholar]
  27. Jessus C., Ducommun B., Beach D. Direct activation of cdc2 with phosphatase: identification of p13suc1-sensitive and insensitive steps. FEBS Lett. 1990 Jun 18;266(1-2):4–8. doi: 10.1016/0014-5793(90)90002-c. [DOI] [PubMed] [Google Scholar]
  28. Kao J. P., Alderton J. M., Tsien R. Y., Steinhardt R. A. Active involvement of Ca2+ in mitotic progression of Swiss 3T3 fibroblasts. J Cell Biol. 1990 Jul;111(1):183–196. doi: 10.1083/jcb.111.1.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kapiloff M. S., Mathis J. M., Nelson C. A., Lin C. R., Rosenfeld M. G. Calcium/calmodulin-dependent protein kinase mediates a pathway for transcriptional regulation. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3710–3714. doi: 10.1073/pnas.88.9.3710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Krek W., Nigg E. A. Differential phosphorylation of vertebrate p34cdc2 kinase at the G1/S and G2/M transitions of the cell cycle: identification of major phosphorylation sites. EMBO J. 1991 Feb;10(2):305–316. doi: 10.1002/j.1460-2075.1991.tb07951.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lamb N. J., Cavadore J. C., Labbe J. C., Maurer R. A., Fernandez A. Inhibition of cAMP-dependent protein kinase plays a key role in the induction of mitosis and nuclear envelope breakdown in mammalian cells. EMBO J. 1991 Jun;10(6):1523–1533. doi: 10.1002/j.1460-2075.1991.tb07672.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Levine H., 3rd, Sahyoun N. E. Characterization of a soluble Mr-30,000 catalytic fragment of the neuronal calmodulin-dependent protein kinase II. Eur J Biochem. 1987 Nov 2;168(3):481–486. doi: 10.1111/j.1432-1033.1987.tb13442.x. [DOI] [PubMed] [Google Scholar]
  33. Lewin B. Driving the cell cycle: M phase kinase, its partners, and substrates. Cell. 1990 Jun 1;61(5):743–752. doi: 10.1016/0092-8674(90)90181-d. [DOI] [PubMed] [Google Scholar]
  34. Lin C. R., Kapiloff M. S., Durgerian S., Tatemoto K., Russo A. F., Hanson P., Schulman H., Rosenfeld M. G. Molecular cloning of a brain-specific calcium/calmodulin-dependent protein kinase. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5962–5966. doi: 10.1073/pnas.84.16.5962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lock R. B., Ross W. E. Inhibition of p34cdc2 kinase activity by etoposide or irradiation as a mechanism of G2 arrest in Chinese hamster ovary cells. Cancer Res. 1990 Jun 15;50(12):3761–3766. [PubMed] [Google Scholar]
  36. MacNicol M., Jefferson A. B., Schulman H. Ca2+/calmodulin kinase is activated by the phosphatidylinositol signaling pathway and becomes Ca2(+)-independent in PC12 cells. J Biol Chem. 1990 Oct 25;265(30):18055–18058. [PubMed] [Google Scholar]
  37. Matsumoto Y., Yasuda H., Mita S., Marunouchi T., Yamada M. Evidence for the involvement of H1 histone phosphorylation in chromosome condensation. Nature. 1980 Mar 13;284(5752):181–183. doi: 10.1038/284181a0. [DOI] [PubMed] [Google Scholar]
  38. Miyakawa T., Oka Y., Tsuchiya E., Fukui S. Saccharomyces cerevisiae protein kinase dependent on Ca2+ and calmodulin. J Bacteriol. 1989 Mar;171(3):1417–1422. doi: 10.1128/jb.171.3.1417-1422.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Morla A. O., Draetta G., Beach D., Wang J. Y. Reversible tyrosine phosphorylation of cdc2: dephosphorylation accompanies activation during entry into mitosis. Cell. 1989 Jul 14;58(1):193–203. doi: 10.1016/0092-8674(89)90415-7. [DOI] [PubMed] [Google Scholar]
  40. Nishimoto T., Ajiro K., Hirata M., Yamashita K., Sekiguchi M. The induction of chromosome condensation in tsBN2, a temperature-sensitive mutant of BHK21, inhibited by the calmodulin antagonist, W-7. Exp Cell Res. 1985 Feb;156(2):351–358. doi: 10.1016/0014-4827(85)90542-7. [DOI] [PubMed] [Google Scholar]
  41. Nurse P. Universal control mechanism regulating onset of M-phase. Nature. 1990 Apr 5;344(6266):503–508. doi: 10.1038/344503a0. [DOI] [PubMed] [Google Scholar]
  42. Ohta Y., Ohba T., Fukunaga K., Miyamoto E. Serum and growth factors rapidly elicit phosphorylation of the Ca2+/calmodulin-dependent protein kinase II in intact quiescent rat 3Y1 cells. J Biol Chem. 1988 Aug 15;263(23):11540–11547. [PubMed] [Google Scholar]
  43. Ohta Y., Ohba T., Miyamoto E. Ca2+/calmodulin-dependent protein kinase II: localization in the interphase nucleus and the mitotic apparatus of mammalian cells. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5341–5345. doi: 10.1073/pnas.87.14.5341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ohya Y., Anraku Y. Functional expression of chicken calmodulin in yeast. Biochem Biophys Res Commun. 1989 Jan 31;158(2):541–547. doi: 10.1016/s0006-291x(89)80083-x. [DOI] [PubMed] [Google Scholar]
  45. Ohya Y., Kawasaki H., Suzuki K., Londesborough J., Anraku Y. Two yeast genes encoding calmodulin-dependent protein kinases. Isolation, sequencing and bacterial expressions of CMK1 and CMK2. J Biol Chem. 1991 Jul 5;266(19):12784–12794. [PubMed] [Google Scholar]
  46. Osmani A. H., McGuire S. L., Osmani S. A. Parallel activation of the NIMA and p34cdc2 cell cycle-regulated protein kinases is required to initiate mitosis in A. nidulans. Cell. 1991 Oct 18;67(2):283–291. doi: 10.1016/0092-8674(91)90180-7. [DOI] [PubMed] [Google Scholar]
  47. Osmani S. A., Pu R. T., Morris N. R. Mitotic induction and maintenance by overexpression of a G2-specific gene that encodes a potential protein kinase. Cell. 1988 Apr 22;53(2):237–244. doi: 10.1016/0092-8674(88)90385-6. [DOI] [PubMed] [Google Scholar]
  48. Paris J., Le Guellec R., Couturier A., Le Guellec K., Omilli F., Camonis J., MacNeill S., Philippe M. Cloning by differential screening of a Xenopus cDNA coding for a protein highly homologous to cdc2. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):1039–1043. doi: 10.1073/pnas.88.3.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Pausch M. H., Kaim D., Kunisawa R., Admon A., Thorner J. Multiple Ca2+/calmodulin-dependent protein kinase genes in a unicellular eukaryote. EMBO J. 1991 Jun;10(6):1511–1522. doi: 10.1002/j.1460-2075.1991.tb07671.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Pearson R. B., Woodgett J. R., Cohen P., Kemp B. E. Substrate specificity of a multifunctional calmodulin-dependent protein kinase. J Biol Chem. 1985 Nov 25;260(27):14471–14476. [PubMed] [Google Scholar]
  51. Poenie M., Alderton J., Tsien R. Y., Steinhardt R. A. Changes of free calcium levels with stages of the cell division cycle. Nature. 1985 May 9;315(6015):147–149. doi: 10.1038/315147a0. [DOI] [PubMed] [Google Scholar]
  52. Qi M., Hamilton B. J., DeFranco D. v-mos oncoproteins affect the nuclear retention and reutilization of glucocorticoid receptors. Mol Endocrinol. 1989 Aug;3(8):1279–1288. doi: 10.1210/mend-3-8-1279. [DOI] [PubMed] [Google Scholar]
  53. Rasmussen C. D., Means A. R. Calmodulin is involved in regulation of cell proliferation. EMBO J. 1987 Dec 20;6(13):3961–3968. doi: 10.1002/j.1460-2075.1987.tb02738.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Rasmussen C. D., Means A. R. Calmodulin is required for cell-cycle progression during G1 and mitosis. EMBO J. 1989 Jan;8(1):73–82. doi: 10.1002/j.1460-2075.1989.tb03350.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Rasmussen C. D., Means A. R. Calmodulin, cell growth and gene expression. Trends Neurosci. 1989 Nov;12(11):433–438. doi: 10.1016/0166-2236(89)90092-1. [DOI] [PubMed] [Google Scholar]
  56. Rasmussen C. D., Means R. L., Lu K. P., May G. S., Means A. R. Characterization and expression of the unique calmodulin gene of Aspergillus nidulans. J Biol Chem. 1990 Aug 15;265(23):13767–13775. [PubMed] [Google Scholar]
  57. Rasmussen C. D., Simmen R. C., MacDougall E. A., Means A. R. Methods for analyzing bovine papilloma virus-based calmodulin expression vectors. Methods Enzymol. 1987;139:642–654. doi: 10.1016/0076-6879(87)39117-7. [DOI] [PubMed] [Google Scholar]
  58. Richardson H. E., Wittenberg C., Cross F., Reed S. I. An essential G1 function for cyclin-like proteins in yeast. Cell. 1989 Dec 22;59(6):1127–1133. doi: 10.1016/0092-8674(89)90768-x. [DOI] [PubMed] [Google Scholar]
  59. Roberge M., Th'ng J., Hamaguchi J., Bradbury E. M. The topoisomerase II inhibitor VM-26 induces marked changes in histone H1 kinase activity, histones H1 and H3 phosphorylation, and chromosome condensation in G2 phase and mitotic BHK cells. J Cell Biol. 1990 Nov;111(5 Pt 1):1753–1762. doi: 10.1083/jcb.111.5.1753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Roskoski R., Jr Assays of protein kinase. Methods Enzymol. 1983;99:3–6. doi: 10.1016/0076-6879(83)99034-1. [DOI] [PubMed] [Google Scholar]
  61. Russell P., Nurse P. Negative regulation of mitosis by wee1+, a gene encoding a protein kinase homolog. Cell. 1987 May 22;49(4):559–567. doi: 10.1016/0092-8674(87)90458-2. [DOI] [PubMed] [Google Scholar]
  62. Russell P., Nurse P. The mitotic inducer nim1+ functions in a regulatory network of protein kinase homologs controlling the initiation of mitosis. Cell. 1987 May 22;49(4):569–576. doi: 10.1016/0092-8674(87)90459-4. [DOI] [PubMed] [Google Scholar]
  63. Samiei M., Daya-Makin M., Clark-Lewis I., Pelech S. L. Platelet-activating factor- and thrombin-induced stimulation of p34cdc2-cyclin histone H1 kinase activity in platelets. J Biol Chem. 1991 Aug 15;266(23):14889–14892. [PubMed] [Google Scholar]
  64. Sasaki Y., Hidaka H. Calmodulin and cell proliferation. Biochem Biophys Res Commun. 1982 Jan 29;104(2):451–456. doi: 10.1016/0006-291x(82)90658-1. [DOI] [PubMed] [Google Scholar]
  65. Schulman H. The multifunctional Ca2+/calmodulin-dependent protein kinase. Adv Second Messenger Phosphoprotein Res. 1988;22:39–112. [PubMed] [Google Scholar]
  66. Sheng M., Thompson M. A., Greenberg M. E. CREB: a Ca(2+)-regulated transcription factor phosphorylated by calmodulin-dependent kinases. Science. 1991 Jun 7;252(5011):1427–1430. doi: 10.1126/science.1646483. [DOI] [PubMed] [Google Scholar]
  67. Surana U., Robitsch H., Price C., Schuster T., Fitch I., Futcher A. B., Nasmyth K. The role of CDC28 and cyclins during mitosis in the budding yeast S. cerevisiae. Cell. 1991 Apr 5;65(1):145–161. doi: 10.1016/0092-8674(91)90416-v. [DOI] [PubMed] [Google Scholar]
  68. Th'ng J. P., Wright P. S., Hamaguchi J., Lee M. G., Norbury C. J., Nurse P., Bradbury E. M. The FT210 cell line is a mouse G2 phase mutant with a temperature-sensitive CDC2 gene product. Cell. 1990 Oct 19;63(2):313–324. doi: 10.1016/0092-8674(90)90164-a. [DOI] [PubMed] [Google Scholar]
  69. Tobimatsu T., Fujisawa H. Tissue-specific expression of four types of rat calmodulin-dependent protein kinase II mRNAs. J Biol Chem. 1989 Oct 25;264(30):17907–17912. [PubMed] [Google Scholar]
  70. Uhler M. D., McKnight G. S. Expression of cDNAs for two isoforms of the catalytic subunit of cAMP-dependent protein kinase. J Biol Chem. 1987 Nov 5;262(31):15202–15207. [PubMed] [Google Scholar]
  71. Ulloa R. M., Torres H. N., Ochatt C. M., Téllez-Iñn M. T. Ca2+ calmodulin-dependent protein kinase activity in the ascomycetes Neurospora crassa. Mol Cell Biochem. 1991 Apr 10;102(2):155–163. doi: 10.1007/BF00234573. [DOI] [PubMed] [Google Scholar]
  72. Waldmann R., Hanson P. I., Schulman H. Multifunctional Ca2+/calmodulin-dependent protein kinase made Ca2+ independent for functional studies. Biochemistry. 1990 Feb 20;29(7):1679–1684. doi: 10.1021/bi00459a002. [DOI] [PubMed] [Google Scholar]
  73. Yamagata Y., Czernik A. J., Greengard P. Active catalytic fragment of Ca2+/calmodulin-dependent protein kinase II. Purification, characterization, and structural analysis. J Biol Chem. 1991 Aug 15;266(23):15391–15397. [PubMed] [Google Scholar]
  74. Yamauchi T., Ohsako S., Deguchi T. Expression and characterization of calmodulin-dependent protein kinase II from cloned cDNAs in Chinese hamster ovary cells. J Biol Chem. 1989 Nov 15;264(32):19108–19116. [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES