Metabolism-based polycyclic aromatic acetylene inhibition of CYP1B1 in 10T1/2 cells potentiates aryl hydrocarbon receptor activity

Toxicology and Applied Pharmacology

Volumn 161, Issue 2, 1999, Pages 123-139

  Alexander, David L ^a   Zhang, Leying ^a   Foroozesh, Maryam ^b   Alworth, William L ^c   Jefcoate, Colin R ^a  

^a UNIVERSITY OF WISCONSIN MADISON   (United States)

^b XAVIER UNIVERSITY OF LOUISIANA   (United States)

^c Tulane University   (United States)

Author keywords

AhR; CYP1B1; Inhibitor; PAH; TCDD

Indexed keywords

2,3,7,8 TETRACHLORODIBENZO PARA DIOXIN; ACETYLENE DERIVATIVE; ALKYNE DERIVATIVE; AROMATIC HYDROCARBON RECEPTOR; CARBON MONOXIDE; CYTOCHROME P450 ISOENZYME; DIMETHYLBENZ[A]ANTHRACENE; MESSENGER RNA; OLIGONUCLEOTIDE; POLYCYCLIC AROMATIC COMPOUND; POLYCYCLIC AROMATIC HYDROCARBON; PYRENE DERIVATIVE; RECEPTOR BLOCKING AGENT; XENOBIOTIC AGENT;

ANIMAL CELL; ARTICLE; CONTROLLED STUDY; ENZYME METABOLISM; FIBROBLAST; HEPATOMA CELL; MOUSE; NONHUMAN; RECEPTOR DOWN REGULATION; TOXICOKINETICS;

ANIMALIA;

EID: 0032804346     PISSN: 0041008X     EISSN: None     Source Type: Journal    
DOI: 10.1006/taap.1999.8794     Document Type: Article

References (92)

1. Alexander D. L., Eltom S. E., Jefcoate C. R. Ah receptor regulation of CYP1B1 expression in primary mouse embryo-derived cells. Cancer Res. 57:1997;4498-4506.
2. Alexander D. L., Ganem L. G., Fernandez-Salguero P., Gonzalez F., Jefcoate C. R. Aryl-hydrocarbon receptor is an inhibitory regulator of lipid synthesis and of commitment to adipogenesis. J. Cell Sci. 111:1998;3311-3322.
3. Alworth W. L., Viaje A., Sandoval A., Warren B. S., Slaga T. J. Potent inhibitory effects of suicide inhibitors of P450 isozymes on 7,12-dimethylbenz[a]anthracene and benzo[a]pyrene initiated skin tumors. Carcinogenesis. 12:1991;1209-1215.
4. Antonsson C., Arulampalam V., Whitelaw M. L., Pettersson S., Poellinger L. Constitutive function of the basic helix-loop-helix/PAS factor Arnt. Regulation of target promoters via the E box motif. J. Biol. Chem. 270:1995a;13968-13972.
5. Antonsson C., Whitelaw M. L., McGuire J., Gustafsson J. A., Poellinger L. Distinct roles of the molecular chaperone hsp90 in modulating dioxin receptor function via the basic helix-loop-helix and PAS domains. Mol. Cell. Biol. 15:1995b;756-765.
6. Bejjani B. A., Lewis R. A., Tomey K. F., Anderson K. L., Dueker D. K., Jabak M., Astle W. F., Otterud B., Leppert M., Lupski J. R. Mutations in CYP1B1, the gene for cytochrome P4501B1, are the predominant cause of primary congenital glaucoma in Saudi Arabia. Am. J. Hum. Genet. 62:1998;325-333.
7. Berghard A., Gradin K., Toftgard R. The stability of dioxin-receptor ligands influences cytochrome P450IA1 expression in human keratinocytes. Carcinogenesis. 13:1992;651-655.
8. Bhat R., Bresnick E. Glycine N-methyltransferase is an example of functional diversity. Role as a polycyclic aromatic hydrocarbon-binding receptor. J. Biol. Chem. 272:1997;21221-21226.
9. Boucher P. D., Piechocki M. P., Hines R. N. Partial characterization of the human CYP1A1 negatively acting transcription factor and mutational analysis of its cognate DNA recognition sequence. Mol. Cell. Biol. 15:1995;5144-5151.
10. Boucher P. D., Ruch R. J., Hines R. N. Specific nuclear protein binding to a negative regulatory element on the human CYP1A1 gene. J. Biol. Chem. 268:1993;17384-17391.
11. 125I]2-iodo-7,8-dibromodibenzo-p-dioxin. Mol. Pharmacol. 34:1988;229-237.
12. Bradfield C. A., Poland A. A competitive binding assay for 2,3,7,8-tetrachlorodibenzo-p-dioxin and related ligands of the Ah receptor. Mol. Pharmacol. 34:1988;682-688.
13. Brake P. B., Jefcoate C. R. Regulation of cytochrome P4501B1 in cultured rat adrenocortical cells by cyclic adenosine 3′,5′-monophosphate and 2,3,7,8-tetrachlorodibenzo-p-dioxin. Endocrinology. 136:1995;5034-5041.
14. Brake P. B., Zhang L., Jefcoate C. R. Aryl hydrocarbon receptor regulation of cytochrome P4501B1 in rat mammary fibroblasts: Evidence for transcriptional repression by glucocorticoids. Mol. Pharmacol. 54:1998;825-833.
15. Buters J. T., Sakai S., Richter T., Pineau T., Alexander D. L., Savas U., Doehmer J., Ward J. M., Jefcoate C. R., Gonzalez F. J. Cytochrome P450 CYP1B1 determines susceptibility to 7,12-dimethylbenz[a]anthracene-induced lymphomas. Proc. Natl. Acad. Sci. USA. 96:1999;1977-1982.
16. Chen Y. H., Tukey R. H. Protein kinase C modulates regulation of the CYP1A1 gene by the aryl hydrocarbon receptor. J. Biol. Chem. 71:1996;26261-26266.
17. Christou M., Keith I. M., Shen X., Schroeder M. E., Jefcoate C. R. Reversal of cytochrome P-4501A1 and P-450-EF expression in MCA-C3H/10T1/2 cell-derived tumors as compared to cultured cells. Cancer Res. 53:1993;968-976.
18. Christou M., Savas U., Schroeder S., Shen X., Thompson T., Gould M. N., Jefcoate C. R. Cytochromes CYP1A1 and CYP1B1 in the rat mammary gland: Cell-specific expression and regulation by polycyclic aromatic hydrocarbons and hormones. Mol. Cell. Endocrinol. 115:1995;41-50.
19. 3H]methylcholanthrene and formation of a 165-kDa complex between the ligand-binding subunit and a novel cytosolic protein. J. Biol. Chem. 269:1994;12129-12136.
20. Eltom S. E., Larsen M. C., Jefcoate C. R. Expression of CYP1B1 but not CYP1A1 by primary cultured human mammary stromal fibroblasts constitutively and in response to dioxin exposure: Role of the Ah receptor. Carcinogenesis. 19:1998;1437-1444.
21. Fernandez-Salguero P., Pineau T., Hilbert D. M., McPhail T., Lee S. S., Kimura S., Nebert D. W., Rudikoff S., Ward J. M., Gonzalez F. J. Immune system impairment and hepatic fibrosis in mice lacking the dioxin-binding Ah receptor [see comments]. Science. 268:1995;722-726.
22. Foroozesh M., Primrose G., Guo Z., Bell L. C., Alworth W. L., Guengerich F. P. Aryl acetylenes as mechanism-based inhibitors of cytochrome P450-dependent monooxygenase enzymes. Chem. Res. Toxicol. 10:1997;91-102.
23. Gasiewicz T. A., Kende A. S., Rucci G., Whitney B., Willey J. J. Analysis of structural requirements for Ah receptor antagonist activity: Ellipticines, flavones, and related compounds. Biochem. Pharmacol. 52:1996;1787-1803.
24. Giannone J. V., Li W., Probst M., Okey A. B. Prolonged depletion of AH receptor without alteration of receptor mRNA levels after treatment of cells in culture with 2,3,7,8-tetrachlorodibenzo-p-dioxin. Biochem. Pharmacol. 55:1998;489-497.
25. Giannone J. V., Okey A. B., Harper P. A. Characterization of polyclonal antibodies to the aromatic hydrocarbon receptor. Can. J. Physiol. Pharmacol. 73:1995;7-17.
26. Gonzalez F. J., Fernandez-Salguero P. The aryl hydrocarbon receptor: Studies using the AhR-null mice. Drug Metab. Dispos. 26:1998;1194-1198.
27. Gradin K., Whitelaw M. L., Toftgard R., Poellinger L., Berghard A. A tyrosine kinase-dependent pathway regulates ligand-dependent activation of the dioxin receptor in human keratinocytes. J. Biol. Chem. 269:1994;23800-23807.
28. Halpert J. R. Structural basis of selective cytochrome P450 inhibition. Annu. Rev. Pharmacol. Toxicol. 35:1995;29-53.
29. Halpert J. R., Guengerich F. P., Bend J. R., Correia M A. Selective inhibitors of cytochromes P450. Toxicol. Appl. Pharmacol. 125:1994;163-175.
30. Hankinson O. The aryl hydrocarbon receptor complex. Annu. Rev. Pharmacol. Toxicol. 35:1995;307-340.
31. Henry E. C., Kende A. S., Rucci G., Totleben M. J., Willey J. J., Dertinger S. D., Pollenz R. S., Jones J. P., Gasiewicz T. A. Flavone antagonists bind competitively with 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD) to the aryl hydrocarbon receptor but inhibit nuclear uptake and transformation. Mol. Pharmacol. 55:1999;716-725.
32. Ho D., Gill K., Fahl W. E. Benz[a]anthracene and 3-methylcholanthrene induction of cytochrome P-450 in C3H/10T1/2 mouse fibroblasts. Modulating role of cytotoxic 3-methylcholanthrene metabolites. Mol. Pharmacol. 23:1983;198-205.
33. Hopkins N. E., Foroozesh M. K., Alworth W. L. Suicide inhibitors of cytochrome P450 1A1 and P450 2B1. Biochem. Pharmacol. 44:1992;787-796.
34. Ikegwuonu F. I., Christou M., Jefcoate C. R. Regulation of cytochrome P4501B1 (CYP1B1) in mouse embryo fibroblast (C3H10T1/2) cells by protein kinase C (PKC). Biochem. Pharmacol. 57:1999;619-630.
35. Israel D. I., Estolano M. G., Galeazzi D. R., Whitlock J. P. Jr. Superinduction of cytochrome P1-450 gene transcription by inhibition of protein synthesis in wild type and variant mouse hepatoma cells. J. Biol. Chem. 260:1985;5648-5653.
36. Janknecht R., Nordheim A. MAP kinase-dependent transcriptional coactivation by Elk-1 and its cofactor CBP. Biochem. Biophys. Res. Commun. 228:1996;831-837.
37. Kallia P. K., Okamoto K., O'Brien S., Carrero P., Makino Y., Tanaka H., Poellinger L. Signal transduction in hypoxix cells: Inducible nuclear translocation and recruitment of the CBP/p300 coactivator by the hypoxia-inducible factor-1α EMBO J. 17:1999;6573-6588.
38. Kobayashi A., Numayama-Tsurta K., Sogawa K., Fuji-Kuriyama Y. CBP/P300 functions as a possible transcriptional coactivator of Ah receptor nuclear translocation (Arnt). J. Biochem. (Tokyo). 122:1997;703-710.
39. Larsen, M. C, Alexander, D. L, Brake, P. B, Angus, W. G. R, Hanlon, P. R, Doehmer, J, Foroozesh, M, Alworth, W. L, and, Jefcoate, C. R. 1999, Mechanism-based aryl acetylenic compounds discriminate between CYP1B1 and CYP1A1 in inhibiting 7,12-dimethylbenz(a)anthracene metabolsim in cultured cells: Species differences in potency and selectivity. Manuscript in preparation.
40. Li S. Y., Dougherty J. J. Inhibitors of serine/threonine-specific protein phosphatases stimulate transcription by the Ah receptor/Arnt dimer by affecting a step subsequent to XRE binding. Arch. Biochem. Biophys. 340:1997;73-82.
41. Lindros K. O., Oinonen T., Johansson I., Ingelman-Sundberg M. Selective centrilobular expression of the aryl hydrocarbon receptor in rat liver. J. Pharmacol. Exp. Ther. 280:1997;506-511.
42. Long W. P., Chen X., Perdew G. H. Protein kinase C modulates aryl hydrocarbon receptor nuclear translocator protein-mediated transactivation potential in a dimer context. J. Biol. Chem. 274:1999;12391-12400.
43. Lu Y. F., Santostefano M., Cunningham B. D., Threadgill M. D., Safe S. Substituted flavones as aryl hydrocarbon (Ah) receptor agonists and antagonists. Biochem. Pharmacol. 51:1996;1077-1087.
44. Lusska A., Shen E., Whitlock J. P. J. Protein-DNA interactions at a dioxin-response element enhancer. Analysis of six bona-fide DNA-binding sites for the liganded Ah receptor. J. Biol. Chem. 268:1993;6575-6580.
45. Lusska A., Wu L., Whitlock J. P. Jr. Superinduction of CYP1A1 transcription by cycloheximide. Role of the DNA binding site for the liganded Ah receptor. J. Biol. Chem. 267:1992;15146-15151.
46. Mahon M. J., Gasiewicz T. A. Ah receptor phosphorylation: Localization of phosphorylation sites to the C-terminal half of the protein. Arch. Biochem. Biophys. 318:1995;166-174.
47. Merchant M., Morrison V., Santostefano M., Safe S. Mechanism of action of aryl hydrocarbon receptor antagonists: Inhibition of 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced CYP1A1 gene expression. Arch. Biochem. Biophys. 298:1992a;389-394.
48. Merchant M., Wang X., Kamps C., Rosengren R., Morrison V., Safe S. Mechanism of benzo[a]pyrene-induced Cyp1a-1 gene expression in mouse Hepa 1c1c7 cells: Role of the nuclear 6 s and 4 s proteins. Arch. Biochem. Biophys. 292:1992b;250-257.
49. Mimura J., Ema M., Sogawa K., Fujii-Kuriyama Y. Identification of a novel mechanism of regulation of Ah (dioxin) receptor function. Genes Dev. 13:1999;20-25.
50. Morgan E. T., Sewer M. B., Iber H., Gonzalez F. J., Lee Y.-H., Tukey R. H., Okino S., Vu T., Chen Y., Sidhu J. S., Omiecinski C. J. Physiological and pathological regulation of cytochrome P450. Drug Metab. Dispos. 26:1998;1232-1240.
51. Nebert D. W. Drug-metabolizing enzymes in ligand-modulated transcription. Biochem. Pharmacol. 47:1994;25-37.
52. Nebert D. W., Petersen D. D., Fornace A. J. Jr. Cellular responses to oxidative stress: The [Ah] gene battery as a paradigm. Environ. Health Perspect. 88:1990;13-25.
53. Norris J. D., Fan D., Stallcup M. R., McDonnell D. P. Enhancement of estrogen receptor transcriptional activity by the coactivator GRIP-1 highlights the role of activation function 2 in determining estrogen receptor pharmacology. J. Biol. Chem. 273:1998;6679-6688.
54. Okey A. B., Riddick D. S., Harper P. A. Molecular biology of the aromatic hydrocarbon (dioxin) receptor. Trends Pharmacol. Sci. 15:1994;226-232.
55. Otto S., Bhattacharyya K. K., Jefcoate C. R. Polycyclic aromatic hydrocarbon metabolism in rat adrenal, ovary, and testis microsomes is catalyzed by the same novel cytochrome P450 (P450RAP). Endocrinology. 131:1992;3067-3076.
56. Parker D., Jhala U. S., Radhakrishnan I., Yaffe M. B., Reyes C., Shulman A. I., Cantley L. C., Wright P. E., Montminy M. Analysis of an activator:coactivator complex reveals an essential role for secondary structure in transcriptional activation. Mol. Cell. 2:1998;353-359.
57. Penning T. M., Ohnishi S. T., Ohnishi T., Harvey R. G. Generation of reactive oxygen species during the enzymatic oxidation of polycyclic aromatic hydrocarbon trans-dihydrodiols catalyzed by dihydrodiol dehydrogenase. Chem. Res. Toxicol. 9:1996;84-92.
58. Perdew G. H., Bradfield C. A. Mapping the 90 kDa heat shock protein binding region of the Ah receptor. Biochem. Mol. Biol. Int. 39:1996;589-593.
59. Pongratz I., Mason G. G., Poellinger L. Dual roles of the 90-kDa heat shock protein hsp90 in modulating functional activities of the dioxin receptor. J. Biol. Chem. 267:1992;13728-13734.
60. Pongratz I., Stromstedt P. E., Mason G. G., Poellinger L. Inhibition of the specific DNA binding activity of the dioxin receptor by phosphatase treatment. J. Biol. Chem. 266:1991;16813-16817.
61. Pottenger L. H., Christou M., Jefcoate C. R. Characterization of a novel cytochrome P-450 from the transformable cell line, C3H10T1/2. Arch. Biochem. Biophys. 286:1990;488-497.
62. Pottenger L. H., Christou M., Jefcoate C. R. Purification and immunological characterization of a novel cytochrome P450 from C3H/10T1/2 cells. Arch. Biochem. Biophys. 286:1991;488-497.
63. 3H]tetrachlorodibenzo-p-dioxin. Biochem. Biophys. Res. Commun. 172:1990;698-704.
64. Prokipcak R. D., Okey A. B. Downregulation of the Ah receptor in mouse hepatoma cells treated in culture with 2,3,7,8-tetrachlorodibenzo-p-dioxin. Can. J. Physiol. Pharmacol. 69:1991;1204-1210.
65. Puga A., Hoffer A., Zhou S., Bohm J. M., Leikauf G. D., Shertzer H. G. Sustained increase in intracellular free calcium and activation of cyclooxygenase-2 expression in mouse hepatoma cells treated with dioxin. Biochem. Pharmacol. 54:1997;1287-1296.
66. Puga A., Raychaudhuri B., Salata K., Zhang Y. H., Nebert D. W. Stable expression of mouse Cyp1a1 and human CYP1A2 cDNAs transfected into mouse hepatoma cells lacking detectable P450 enzyme activity. DNA Cell Biol. 9:1990;425-436.
67. RayChaudhuri B., Nebert D. W., Puga A. The murine Cyp1a-1 gene negatively regulates its own transcription and that of other members of the aromatic hydrocarbon-responsive [Ah] gene battery. Mol. Endocrinol. 4:1990;1773-1781.
68. Reick M., Robertson R. W., Pasco D. S., Fagan J. B. Down-regulation of nuclear aryl hydrocarbon receptor DNA-binding and transactivation functions: Requirement for a labile or inducible factor. Mol. Cell. Biol. 14:1994;5653-5660.
69. Reiners J. J. Jr., Jones C. L., Hong N., Clift R. E., Elferink C. Downregulation of aryl hydrocarbon receptor function and cytochrome P450 1A1 induction by expression of Ha-ras oncogenes. Mol. Carcinogen. 19:1997;91-100.
70. Riddick D. S., Huang Y., Harper P. A., Okey A. B. 2,3,7,8-Tetrachlorodibenzo-p-dioxin versus 3-methylcholanthrene: Comparative studies of Ah receptor binding, transformation, and induction of CYP1A1. J. Biol. Chem. 269:1994;12118-12128.
71. Roberts E. S., Hopkins N. E., Foroozesh M., Alworth W. L., Halpert J. R., Hollenberg P. F. Inactivation of cytochrome P450s 2B1, 2B4, 2B6, and 2B11 by arylalkynes. Drug Metab. Dispos. 25:1997;1242-1248.
72. Rowlands J. C., Gustafsson J. A. Aryl hydrocarbon receptor-mediated signal transduction. Crit. Rev. Toxicol. 27:1997;109-134.
73. Sambrook J., Fritsch E. F., Maniatis T. Molecular Cloning: A Laboratory Manual. 1989;Cold Spring Harbor Laboratory Press, Plainview.
74. Savas U., Bhattacharyya K. K., Christou M., Alexander D. L., Jefcoate C. R. Mouse cytochrome P-450EF, representative of a new 1B subfamily of cytochrome P-450s. Cloning, sequence determination, and tissue expression. J. Biol. Chem. 269:1994;14905-14911.
75. Savas U., Carstens C. P., Jefcoate C. R. Biological oxidations and P450 reactions. Recombinant mouse CYP1B1 expressed in Escherichia coli exhibits selective binding by polycyclic hydrocarbons and metabolism which parallels C3H10T1/2 cell microsomes, but differs from human recombinant CYP1B1. Arch. Biochem. Biophys. 347:1997;181-192.
76. Savas U., Jefcoate C. R. Dual regulation of cytochrome P450EF expression via the aryl hydrocarbon receptor and protein stabilization in C3H/10T1/2 cells. Mol. Pharmacol. 45:1994;1153-1159.
77. Schmidt J. V., Bradfield C. A. Ah receptor signaling pathways. Annu. Rev. Cell Dev. Biol. 12:1996;55-89.
78. Shimada T., Hayes C. L., Yamazaki H., Amin S., Hecht S. S., Guengerich F. P., Sutter T. R. Activation of chemically diverse procarcinogens by human cytochrome P-450 1B1. Cancer Res. 56:1996;2979-2984.
79. Shimada T., Yamazaki H., Foroozesh M., Hopkins N. E., Alworth W. L., Guengerich F. P. Selectivity of polycyclic inhibitors for human cytochrome P450s 1A1, 1A1, and 1B1. Chem. Res. Toxicol. 11:1998;1043-1059.
80. Silverman R. B., Hiebert C. K. Inactivation of monoamine oxidase A by the monoamine oxidase B inactivators 1-phenylcyclopropylamine, 1-benzylcyclopropylamine, and N-cyclopropyl-alpha-methylbenzylamine. Biochemistry. 27:1988;8448-8453.
81. Spink D. C., Spink B. C., Cao J. Q., DePasquale J. A., Pentecost B. T., Fasco M. J., Li Y., Sutter T. R. Differential expression of CYP1A1 and CYP1B1 in human breast epithelial cells and breast tumor cells. Carcinogenesis. 19:1998;291-298.
82. Swanson H. I., Yang J. H. The aryl hydrocarbon receptor interacts with transcription factor IIB. Mol. Pharmacol. 54:1998;671-677.
83. Tang Y. M., Wo Y. Y. P., Stewart J., Hawkins A. L., Griffin C. A., Sutter T. R., Greenlee W. F. Isolation and characterization of the human cytochrome P450 CYP1B1 gene. J. Biol. Chem. 271:1996;28324-28330.
84. Tannheimer S. L., Lauer F. T., Lane J., Burchiel S. W. Factors influencing elevation of intracellular Ca2+ in the MCF-10A human mammary epithelial cell line by carcinogenic polycyclic aromatic hydrocarbons. Mol. Carcinog. 25:1999;48-54.
85. Tian Y., Ke S., Denison M. S., Rabson A. B., Gallo M. A. Ah receptor and NF-kappaB interactions, a potential mechanism for dioxin toxicity. J. Biol. Chem. 274:1999;510-515.
86. Vasiliou V., Puga A., Nebert D. W. Negative regulation of the murine cytosolic aldehyde dehydrogenase-3 (Aldh-3c) gene by functional CYP1A1 and CYP1A2 proteins. Biochem. Biophys. Res. Commun. 187:1992;413-419.
87. Vasiliou V., Theurer M. J., Puga A., Reuter S. F., Nebert D. W. Mouse dioxin-inducible NAD(P)H: menadione oxidoreductase: NMO1 cDNA sequence and genetic differences in mRNA levels. Pharmacogenetics. 4:1994;341-348.
88. Waller C. L., McKinney J. D. Three-dimensional quantitative structure-activity relationships of dioxins and dioxin-like compounds: Model validation and Ah receptor characterization. Chem. Res. Toxicol. 8:1995;847-858.
89. Wasserman W. W., Fahl W. E. Functional antioxidant responsive elements. Proc. Natl. Acad. Sci. USA. 94:1997;5361-5366.
90. Whitlock J. P. Jr. Mechanistic aspects of dioxin action. Chem. Res. Toxicol. 6:1993;754-763.
91. Whitlock J. P. Jr., Chichester C. H., Bedgood R. M., Okino S. T., Ko H. P., Ma Q., Dong L., Li H., Clarke-Katzenberg R. Induction of drug-metabolizing enzymes by dioxin. Drug Metab. Rev. 29:1997;1107-1127.
92. Zhang L., Savas U., Alexander D. L., Jefcoate C. R. Characterization of the mouse Cyp1B1 gene. Identification of an enhancer region that directs aryl hydrocarbon receptor-mediated constitutive and induced expression. J. Biol. Chem. 273:1998;5174-5183.