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. 1996 Jul;16(7):3901–3908. doi: 10.1128/mcb.16.7.3901

Tumor cell complementation groups based on myogenic potential: evidence for inactivation of loci required for basic helix-loop-helix protein activity.

A N Gerber 1, S J Tapscott 1
PMCID: PMC231387  PMID: 8668208

Abstract

Basic helix-loop-helix (bHLH) proteins mediate terminal differentiation in many lineages. By using the bHLH protein MyoD, which can dominantly activate the myogenic differentiation program in numerous cell types, we demonstrated that recessive defects in bHLH protein function are present in human tumor lines. In contrast to prior work with primary cell cultures, MyoD did not activate the myogenic program in six of the eight tumor lines we tested. Cell fusions between the MyoD-defective lines and fibroblasts restored MyoD activity, indicating that the deficiency of a gene or factor prevents bHLH protein function in the tumor lines. Fusions between certain pairings of the MyoD-defective lines also restored MyoD activity, allowing the tumor lines to be assigned to complementation groups on the basis of their ability to execute the myogenic program and indicating that multiple mechanisms exist for abrogation of bHLH protein activity. These groups provide a basis for identifying genes critical for bHLH-mediated differentiation and tumor progression by using genetic complementation.

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Selected References

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  1. Airey J. A., Baring M. D., Sutko J. L. Ryanodine receptor protein is expressed during differentiation in the muscle cell lines BC3H1 and C2C12. Dev Biol. 1991 Nov;148(1):365–374. doi: 10.1016/0012-1606(91)90344-3. [DOI] [PubMed] [Google Scholar]
  2. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  3. Azar C. G., Scavarda N. J., Reynolds C. P., Brodeur G. M. Multiple defects of the nerve growth factor receptor in human neuroblastomas. Cell Growth Differ. 1990 Sep;1(9):421–428. [PubMed] [Google Scholar]
  4. Bader S. A., Fasching C., Brodeur G. M., Stanbridge E. J. Dissociation of suppression of tumorigenicity and differentiation in vitro effected by transfer of single human chromosomes into human neuroblastoma cells. Cell Growth Differ. 1991 May;2(5):245–255. [PubMed] [Google Scholar]
  5. Bengal E., Ransone L., Scharfmann R., Dwarki V. J., Tapscott S. J., Weintraub H., Verma I. M. Functional antagonism between c-Jun and MyoD proteins: a direct physical association. Cell. 1992 Feb 7;68(3):507–519. doi: 10.1016/0092-8674(92)90187-h. [DOI] [PubMed] [Google Scholar]
  6. Blanar M. A., Crossley P. H., Peters K. G., Steingrímsson E., Copeland N. G., Jenkins N. A., Martin G. R., Rutter W. J. Meso1, a basic-helix-loop-helix protein involved in mammalian presomitic mesoderm development. Proc Natl Acad Sci U S A. 1995 Jun 20;92(13):5870–5874. doi: 10.1073/pnas.92.13.5870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brennan T. J., Edmondson D. G., Li L., Olson E. N. Transforming growth factor beta represses the actions of myogenin through a mechanism independent of DNA binding. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3822–3826. doi: 10.1073/pnas.88.9.3822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Carreau M., Eveno E., Quilliet X., Chevalier-Lagente O., Benoit A., Tanganelli B., Stefanini M., Vermeulen W., Hoeijmakers J. H., Sarasin A. Development of a new easy complementation assay for DNA repair deficient human syndromes using cloned repair genes. Carcinogenesis. 1995 May;16(5):1003–1009. doi: 10.1093/carcin/16.5.1003. [DOI] [PubMed] [Google Scholar]
  9. Carriaga M. T., Henson D. E. The histologic grading of cancer. Cancer. 1995 Jan 1;75(1 Suppl):406–421. doi: 10.1002/1097-0142(19950101)75:1+<406::aid-cncr2820751322>3.0.co;2-w. [DOI] [PubMed] [Google Scholar]
  10. Cerosaletti K. M., Shapero M. H., Fournier R. E. Cloning mammary cell cDNAs from 17q12-q23 using interspecific somatic cell hybrids and subtractive hybridization. Genomics. 1995 Jan 1;25(1):226–237. doi: 10.1016/0888-7543(95)80130-e. [DOI] [PubMed] [Google Scholar]
  11. Chen P., Ellmore N., Weissman B. E. Functional evidence for a second tumor suppressor gene on human chromosome 17. Mol Cell Biol. 1994 Jan;14(1):534–542. doi: 10.1128/mcb.14.1.534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Choi J., Costa M. L., Mermelstein C. S., Chagas C., Holtzer S., Holtzer H. MyoD converts primary dermal fibroblasts, chondroblasts, smooth muscle, and retinal pigmented epithelial cells into striated mononucleated myoblasts and multinucleated myotubes. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7988–7992. doi: 10.1073/pnas.87.20.7988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Clark J., Rocques P. J., Braun T., Bober E., Arnold H. H., Fisher C., Fletcher C., Brown K., Gusterson B. A., Carter R. L. Expression of members of the myf gene family in human rhabdomyosarcomas. Br J Cancer. 1991 Dec;64(6):1039–1042. doi: 10.1038/bjc.1991.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Cserjesi P., Brown D., Ligon K. L., Lyons G. E., Copeland N. G., Gilbert D. J., Jenkins N. A., Olson E. N. Scleraxis: a basic helix-loop-helix protein that prefigures skeletal formation during mouse embryogenesis. Development. 1995 Apr;121(4):1099–1110. doi: 10.1242/dev.121.4.1099. [DOI] [PubMed] [Google Scholar]
  15. Davis R. L., Weintraub H. Acquisition of myogenic specificity by replacement of three amino acid residues from MyoD into E12. Science. 1992 May 15;256(5059):1027–1030. doi: 10.1126/science.1317057. [DOI] [PubMed] [Google Scholar]
  16. Dienstman S. R., Holtzer H. Skeletal myogenesis. Control of proliferation in a normal cell lineage. Exp Cell Res. 1977 Jul;107(2):355–364. doi: 10.1016/0014-4827(77)90357-3. [DOI] [PubMed] [Google Scholar]
  17. Felsenfeld A. L., Curry M., Kimmel C. B. The fub-1 mutation blocks initial myofibril formation in zebrafish muscle pioneer cells. Dev Biol. 1991 Nov;148(1):23–30. doi: 10.1016/0012-1606(91)90314-s. [DOI] [PubMed] [Google Scholar]
  18. Giangrande A. Proneural genes influence gliogenesis in Drosophila. Development. 1995 Feb;121(2):429–438. doi: 10.1242/dev.121.2.429. [DOI] [PubMed] [Google Scholar]
  19. Goyette M. C., Cho K., Fasching C. L., Levy D. B., Kinzler K. W., Paraskeva C., Vogelstein B., Stanbridge E. J. Progression of colorectal cancer is associated with multiple tumor suppressor gene defects but inhibition of tumorigenicity is accomplished by correction of any single defect via chromosome transfer. Mol Cell Biol. 1992 Mar;12(3):1387–1395. doi: 10.1128/mcb.12.3.1387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Graña X., Reddy E. P. Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Oncogene. 1995 Jul 20;11(2):211–219. [PubMed] [Google Scholar]
  21. Gu W., Schneider J. W., Condorelli G., Kaushal S., Mahdavi V., Nadal-Ginard B. Interaction of myogenic factors and the retinoblastoma protein mediates muscle cell commitment and differentiation. Cell. 1993 Feb 12;72(3):309–324. doi: 10.1016/0092-8674(93)90110-c. [DOI] [PubMed] [Google Scholar]
  22. Guillemot F., Lo L. C., Johnson J. E., Auerbach A., Anderson D. J., Joyner A. L. Mammalian achaete-scute homolog 1 is required for the early development of olfactory and autonomic neurons. Cell. 1993 Nov 5;75(3):463–476. doi: 10.1016/0092-8674(93)90381-y. [DOI] [PubMed] [Google Scholar]
  23. Guillemot F., Nagy A., Auerbach A., Rossant J., Joyner A. L. Essential role of Mash-2 in extraembryonic development. Nature. 1994 Sep 22;371(6495):333–336. doi: 10.1038/371333a0. [DOI] [PubMed] [Google Scholar]
  24. Habraken Y., Sung P., Prakash L., Prakash S. Human xeroderma pigmentosum group G gene encodes a DNA endonuclease. Nucleic Acids Res. 1994 Aug 25;22(16):3312–3316. doi: 10.1093/nar/22.16.3312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Halevy O., Novitch B. G., Spicer D. B., Skapek S. X., Rhee J., Hannon G. J., Beach D., Lassar A. B. Correlation of terminal cell cycle arrest of skeletal muscle with induction of p21 by MyoD. Science. 1995 Feb 17;267(5200):1018–1021. doi: 10.1126/science.7863327. [DOI] [PubMed] [Google Scholar]
  26. He J., Allen J. R., Collins V. P., Allalunis-Turner M. J., Godbout R., Day R. S., 3rd, James C. D. CDK4 amplification is an alternative mechanism to p16 gene homozygous deletion in glioma cell lines. Cancer Res. 1994 Nov 15;54(22):5804–5807. [PubMed] [Google Scholar]
  27. Hock R. A., Miller A. D. Retrovirus-mediated transfer and expression of drug resistance genes in human haematopoietic progenitor cells. Nature. 1986 Mar 20;320(6059):275–277. doi: 10.1038/320275a0. [DOI] [PubMed] [Google Scholar]
  28. Hong L. L., Johannsen L., Krueger J. M. Modulation of human leukocyte antigenDR expression in glioblastoma cells by interferon gamma and other cytokines. J Neuroimmunol. 1991 Dec;35(1-3):139–152. doi: 10.1016/0165-5728(91)90169-8. [DOI] [PubMed] [Google Scholar]
  29. Hosoi H., Sugimoto T., Hayashi Y., Inaba T., Horii Y., Morioka H., Fushiki S., Hamazaki M., Sawada T. Differential expression of myogenic regulatory genes, MyoD1 and myogenin, in human rhabdomyosarcoma sublines. Int J Cancer. 1992 Apr 1;50(6):977–983. doi: 10.1002/ijc.2910500626. [DOI] [PubMed] [Google Scholar]
  30. Höll T., Kleihues P., Yasargil M. G., Wiestler O. D. Cerebellar medullomyoblastoma with advanced neuronal differentiation and hamartomatous component. Acta Neuropathol. 1991;82(5):408–413. doi: 10.1007/BF00296553. [DOI] [PubMed] [Google Scholar]
  31. Jarman A. P., Sun Y., Jan L. Y., Jan Y. N. Role of the proneural gene, atonal, in formation of Drosophila chordotonal organs and photoreceptors. Development. 1995 Jul;121(7):2019–2030. doi: 10.1242/dev.121.7.2019. [DOI] [PubMed] [Google Scholar]
  32. Jaspers N. G., Bootsma D. Genetic heterogeneity in ataxia-telangiectasia studied by cell fusion. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2641–2644. doi: 10.1073/pnas.79.8.2641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kamb A., Gruis N. A., Weaver-Feldhaus J., Liu Q., Harshman K., Tavtigian S. V., Stockert E., Day R. S., 3rd, Johnson B. E., Skolnick M. H. A cell cycle regulator potentially involved in genesis of many tumor types. Science. 1994 Apr 15;264(5157):436–440. doi: 10.1126/science.8153634. [DOI] [PubMed] [Google Scholar]
  34. Kaur G. P., Athwal R. S. Complementation of DNA repair defect in xeroderma pigmentosum cells of group C by the transfer of human chromosome 5. Somat Cell Mol Genet. 1993 Jan;19(1):83–93. doi: 10.1007/BF01233957. [DOI] [PubMed] [Google Scholar]
  35. Kraemer K. H., Coon H. G., Petinga R. A., Barrett S. F., Rahe A. E., Robbins J. H. National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014, USA. Proc Natl Acad Sci U S A. 1975 Jan;72(1):59–63. doi: 10.1073/pnas.72.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lee J. E., Hollenberg S. M., Snider L., Turner D. L., Lipnick N., Weintraub H. Conversion of Xenopus ectoderm into neurons by NeuroD, a basic helix-loop-helix protein. Science. 1995 May 12;268(5212):836–844. doi: 10.1126/science.7754368. [DOI] [PubMed] [Google Scholar]
  37. Li L., Chambard J. C., Karin M., Olson E. N. Fos and Jun repress transcriptional activation by myogenin and MyoD: the amino terminus of Jun can mediate repression. Genes Dev. 1992 Apr;6(4):676–689. doi: 10.1101/gad.6.4.676. [DOI] [PubMed] [Google Scholar]
  38. Li L., Zhou J., James G., Heller-Harrison R., Czech M. P., Olson E. N. FGF inactivates myogenic helix-loop-helix proteins through phosphorylation of a conserved protein kinase C site in their DNA-binding domains. Cell. 1992 Dec 24;71(7):1181–1194. doi: 10.1016/s0092-8674(05)80066-2. [DOI] [PubMed] [Google Scholar]
  39. Lukas J., Parry D., Aagaard L., Mann D. J., Bartkova J., Strauss M., Peters G., Bartek J. Retinoblastoma-protein-dependent cell-cycle inhibition by the tumour suppressor p16. Nature. 1995 Jun 8;375(6531):503–506. doi: 10.1038/375503a0. [DOI] [PubMed] [Google Scholar]
  40. Miller A. D., Rosman G. J. Improved retroviral vectors for gene transfer and expression. Biotechniques. 1989 Oct;7(9):980-2, 984-6, 989-90. [PMC free article] [PubMed] [Google Scholar]
  41. Miner J. H., Wold B. J. c-myc inhibition of MyoD and myogenin-initiated myogenic differentiation. Mol Cell Biol. 1991 May;11(5):2842–2851. doi: 10.1128/mcb.11.5.2842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Molkentin J. D., Black B. L., Martin J. F., Olson E. N. Cooperative activation of muscle gene expression by MEF2 and myogenic bHLH proteins. Cell. 1995 Dec 29;83(7):1125–1136. doi: 10.1016/0092-8674(95)90139-6. [DOI] [PubMed] [Google Scholar]
  43. Neuhold L. A., Wold B. HLH forced dimers: tethering MyoD to E47 generates a dominant positive myogenic factor insulated from negative regulation by Id. Cell. 1993 Sep 24;74(6):1033–1042. doi: 10.1016/0092-8674(93)90725-6. [DOI] [PubMed] [Google Scholar]
  44. Ogawa S., Hangaishi A., Miyawaki S., Hirosawa S., Miura Y., Takeyama K., Kamada N., Ohtake S., Uike N., Shimazaki C. Loss of the cyclin-dependent kinase 4-inhibitor (p16; MTS1) gene is frequent in and highly specific to lymphoid tumors in primary human hematopoietic malignancies. Blood. 1995 Aug 15;86(4):1548–1556. [PubMed] [Google Scholar]
  45. Peng H. Q., Bailey D., Bronson D., Goss P. E., Hogg D. Loss of heterozygosity of tumor suppressor genes in testis cancer. Cancer Res. 1995 Jul 1;55(13):2871–2875. [PubMed] [Google Scholar]
  46. Pereira-Smith O. M., Smith J. R. Genetic analysis of indefinite division in human cells: identification of four complementation groups. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6042–6046. doi: 10.1073/pnas.85.16.6042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Prados J., Melguizo C., Fernandez J. E., Aranega A. E., Alvarez L., Aranega A. Actin, tropomyosin and alpha-actinin as markers of differentiation in human rhabdomyosarcoma cell lines induced with dimethyl sulfoxide. Cell Mol Biol (Noisy-le-grand) 1993 Jul;39(5):525–536. [PubMed] [Google Scholar]
  48. Raza A., Yousuf N., Bokhari S. A., Mehdi A., Masterson M., Lampkin B., Yanik G., Mazewski C., Khan S., Preisler H. Contribution of in vivo proliferation/differentiation studies toward the development of a combined functional and morphologic system of classification of neoplastic diseases. Cancer. 1992 Mar 15;69(6 Suppl):1557–1566. doi: 10.1002/1097-0142(19920315)69:6+<1557::aid-cncr2820691309>3.0.co;2-5. [DOI] [PubMed] [Google Scholar]
  49. Sancho S., Mongini T., Tanji K., Tapscott S. J., Walker W. F., Weintraub H., Miller A. D., Miranda A. F. Analysis of dystrophin expression after activation of myogenesis in amniocytes, chorionic-villus cells, and fibroblasts. A new method for diagnosing Duchenne's muscular dystrophy. N Engl J Med. 1993 Sep 23;329(13):915–920. doi: 10.1056/NEJM199309233291303. [DOI] [PubMed] [Google Scholar]
  50. Scherly D., Nouspikel T., Corlet J., Ucla C., Bairoch A., Clarkson S. G. Complementation of the DNA repair defect in xeroderma pigmentosum group G cells by a human cDNA related to yeast RAD2. Nature. 1993 May 13;363(6425):182–185. doi: 10.1038/363182a0. [DOI] [PubMed] [Google Scholar]
  51. Schiffer D., Giordana M. T., Pezzotta S., Pezzulo T., Vigliani M. C. Medullomyoblastoma: report of two cases. Childs Nerv Syst. 1992 Aug;8(5):268–272. doi: 10.1007/BF00300794. [DOI] [PubMed] [Google Scholar]
  52. Schmidt E. E., Ichimura K., Reifenberger G., Collins V. P. CDKN2 (p16/MTS1) gene deletion or CDK4 amplification occurs in the majority of glioblastomas. Cancer Res. 1994 Dec 15;54(24):6321–6324. [PubMed] [Google Scholar]
  53. Sherr C. J., Roberts J. M. Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev. 1995 May 15;9(10):1149–1163. doi: 10.1101/gad.9.10.1149. [DOI] [PubMed] [Google Scholar]
  54. Silber J. R., Bobola M. S., Ewers T. G., Muramoto M., Berger M. S. O6-alkylguanine DNA-alkyltransferase is not a major determinant of sensitivity to 1,3-bis(2-chloroethyl)-1-nitrosourea in four medulloblastoma cell lines. Oncol Res. 1992;4(6):241–248. [PubMed] [Google Scholar]
  55. Silbergeld D. L., Ali-Osman F., Winn H. R. Induction of transformational changes in normal endothelial cells by cultured human astrocytoma cells. J Neurosurg. 1991 Oct;75(4):604–612. doi: 10.3171/jns.1991.75.4.0604. [DOI] [PubMed] [Google Scholar]
  56. Skapek S. X., Rhee J., Spicer D. B., Lassar A. B. Inhibition of myogenic differentiation in proliferating myoblasts by cyclin D1-dependent kinase. Science. 1995 Feb 17;267(5200):1022–1024. doi: 10.1126/science.7863328. [DOI] [PubMed] [Google Scholar]
  57. Sorensen P. H., Shimada H., Liu X. F., Lim J. F., Thomas G., Triche T. J. Biphenotypic sarcomas with myogenic and neural differentiation express the Ewing's sarcoma EWS/FLI1 fusion gene. Cancer Res. 1995 Mar 15;55(6):1385–1392. [PubMed] [Google Scholar]
  58. Sperduto P. W., Song C. W., Levitt S. H. The in vitro radiosensitization of human glioblastoma with pentoxifylline. Am J Clin Oncol. 1993 Oct;16(5):407–411. doi: 10.1097/00000421-199310000-00007. [DOI] [PubMed] [Google Scholar]
  59. Stegmaier K., Pendse S., Barker G. F., Bray-Ward P., Ward D. C., Montgomery K. T., Krauter K. S., Reynolds C., Sklar J., Donnelly M. Frequent loss of heterozygosity at the TEL gene locus in acute lymphoblastic leukemia of childhood. Blood. 1995 Jul 1;86(1):38–44. [PubMed] [Google Scholar]
  60. Su H. Y., Bos T. J., Monteclaro F. S., Vogt P. K. Jun inhibits myogenic differentiation. Oncogene. 1991 Oct;6(10):1759–1766. [PubMed] [Google Scholar]
  61. Tapscott S. J., Thayer M. J., Weintraub H. Deficiency in rhabdomyosarcomas of a factor required for MyoD activity and myogenesis. Science. 1993 Mar 5;259(5100):1450–1453. doi: 10.1126/science.8383879. [DOI] [PubMed] [Google Scholar]
  62. Vaidya T. B., Weyman C. M., Teegarden D., Ashendel C. L., Taparowsky E. J. Inhibition of myogenesis by the H-ras oncogene: implication of a role for protein kinase C. J Cell Biol. 1991 Aug;114(4):809–820. doi: 10.1083/jcb.114.4.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Weeda G., Wiegant J., van der Ploeg M., Geurts van Kessel A. H., van der Eb A. J., Hoeijmakers J. H. Localization of the xeroderma pigmentosum group B-correcting gene ERCC3 to human chromosome 2q21. Genomics. 1991 Aug;10(4):1035–1040. doi: 10.1016/0888-7543(91)90195-k. [DOI] [PubMed] [Google Scholar]
  64. Weintraub H., Dwarki V. J., Verma I., Davis R., Hollenberg S., Snider L., Lassar A., Tapscott S. J. Muscle-specific transcriptional activation by MyoD. Genes Dev. 1991 Aug;5(8):1377–1386. doi: 10.1101/gad.5.8.1377. [DOI] [PubMed] [Google Scholar]
  65. Weintraub H., Hauschka S., Tapscott S. J. The MCK enhancer contains a p53 responsive element. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4570–4571. doi: 10.1073/pnas.88.11.4570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Weintraub H., Tapscott S. J., Davis R. L., Thayer M. J., Adam M. A., Lassar A. B., Miller A. D. Activation of muscle-specific genes in pigment, nerve, fat, liver, and fibroblast cell lines by forced expression of MyoD. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5434–5438. doi: 10.1073/pnas.86.14.5434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Weintraub H. The MyoD family and myogenesis: redundancy, networks, and thresholds. Cell. 1993 Dec 31;75(7):1241–1244. doi: 10.1016/0092-8674(93)90610-3. [DOI] [PubMed] [Google Scholar]
  68. Xiao S., Li D., Corson J. M., Vijg J., Fletcher J. A. Codeletion of p15 and p16 genes in primary non-small cell lung carcinoma. Cancer Res. 1995 Jul 15;55(14):2968–2971. [PubMed] [Google Scholar]
  69. Zakrzewski S., Sperling K. Genetic heterogeneity of Fanconi's anemia demonstrated by somatic cell hybrids. Hum Genet. 1980;56(1):81–84. doi: 10.1007/BF00281573. [DOI] [PubMed] [Google Scholar]

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