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Abstract:

The E6 oncoproteins of high-risk HPV types 16 and 18 are involved in the development of cervical cancer. Besides its determinant role in carcinogenic progression, HPV E6 oncoprotein has also been instrumental in elucidating fundamental aspects of p53 function and its ubiquitin-proteasome degradation, with counterpart activities in various DNA tumor viruses. Establishing the conformational state and cellular distribution unequivocally for the endogenous protein in HPV-transformed cell lines derived from carcinomas is essential for understanding the underlying mechanism. Recombinant E6 from high-risk strains 16 and 18 folds into soluble oligomers of ∼1.2 MDa, which are thermostable and display cooperative loss of tertiary and secondary structure upon chemical denaturation. Antibodies raised against these assemblies locate E6 evenly distributed in the cells. By depleting the polyclonal serum by immunoblocking with monomeric E6, the nuclei of Hela and CaSki cells become completely devoid of label, indicating that monomeric species are mainly localized in the nucleus and that both monomers and oligomers share epitopes. The monomeric species promote degradation of p53 by the proteasome, which correlates with the nuclear localization we describe. In contrast, the oligomeric E6 does not promote p53 degradation, in agreement with its cytoplasmic localization inferred from the immunoneutralization experiments. Our results indicate that the cytoplasmic species contain conformational epitopes that may arise from yet undefined homo or hetero-oligomers, but its localization otherwise agrees with that of the other group of major E6 targets, those involving PDZ binding domains, which requires further investigation. © 2007 American Chemical Society.

Registro:

Documento: Artículo
Título:High-risk HPV E6 oncoproteins assemble into large oligomers that allow localization of endogenous species in prototypic HPV-transformed cell lines
Autor:García-Alai, M.M.; Dantur, K.I.; Smal, C.; Pietrasanta, L.; De Prat-Gay, G.
Filiación:Instituto Leloir and CONICET, Patricias Argentinas 435, 1405 Buenos Aires, Argentina
Facultad de Ciencias Exactas Y Naturales, UBA, Buenos Aires, Argentina
Palabras clave:Antibodies; Biodegradation; Cells; Monomers; Oligomers; Oncology; Carcinogenic progression; Cervical cancer; Endogenous protein; Ubiquitin-proteasome degradation; Proteins; oligomer; PDZ protein; proteasome; protein antibody; protein E6; protein p53; recombinant protein; ubiquitin; article; cancer growth; carcinogenesis; cell line; cell nucleus; cellular distribution; controlled study; DNA tumor virus; embryo; HeLa cell; human; human cell; nonhuman; particle size; priority journal; protein analysis; protein assembly; protein binding; protein conformation; protein degradation; protein denaturation; protein domain; protein function; protein localization; protein quaternary structure; protein targeting; protein tertiary structure; protein transport; serum; structural proteomics; thermostability; uterine cervix cancer; virus cell transformation; virus strain; virus typing; Wart virus; Base Sequence; Cell Line, Transformed; Cell Transformation, Neoplastic; Cell Transformation, Viral; DNA, Viral; DNA-Binding Proteins; Female; Human papillomavirus 16; Human papillomavirus 18; Humans; Models, Biological; Multiprotein Complexes; Oncogene Proteins, Viral; Protein Folding; Protein Structure, Quaternary; Repressor Proteins; Tumor Suppressor Protein p53; Uterine Cervical Neoplasms; Human papillomavirus
Año:2007
Volumen:46
Número:2
Página de inicio:341
Página de fin:349
DOI: http://dx.doi.org/10.1021/bi602457q
Título revista:Biochemistry
Título revista abreviado:Biochemistry
ISSN:00062960
CODEN:BICHA
CAS:proteasome, 140879-24-9; ubiquitin, 60267-61-0; DNA, Viral; DNA-Binding Proteins; E6 protein, Human papillomavirus type 16; E6 protein, Human papillomavirus type 18; Multiprotein Complexes; Oncogene Proteins, Viral; Repressor Proteins; TP53 protein, human; Tumor Suppressor Protein p53
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v46_n2_p341_GarciaAlai

Referencias:

  • Lowy, D.R., Howley, P.M., (2001) Fields Virology, pp. 2213-2219. , Fields, B. N, Howley, P. M, Griffin, D. E, Lamb, R. A, Martin, M. A, Roizman, B, Straus, S. E, and Knipe, D. M, Eds, pp, Lippincott Williams and Wilkins, Philadelphia
  • Thomas, M., Pim, D., and Banks, L. (2006) in Papillomavirus Research (Campo, M. S., Ed.) pp 115-131, Caister Academic Press, Wymondham, Norfolk, U.K; Helt, A.M., Galloway, D.A., Mechanisms by which DNA tumor virus oncoproteins target the Rb family of pocket proteins (2003) Carcinogenesis, 24, pp. 159-169
  • Zambetti, G.P., (2005) Protein Reviews, pp. 1-23. , Atassi, M. Z, Ed, pp, Springer Science, New York
  • Munger, K., Baldwin, A., Edwards, K.M., Hayakawa, H., Nguyen, C.L., Owens, M., Grace, M., Huh, K., Mechanisms of human papillomavirus-induced oncogenesis (2004) J. Virol, 78, pp. 11451-11460
  • Munger, K., Howley, P.M., Human papillomavirus immortalization and transformation functions (2002) Virus Res, 89, pp. 213-228
  • Durst, M., Gissmann, L., Ikenberg, H., zur Hausen, H., A papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographic regions (1983) Proc. Natl. Acad. Sci. U.S.A, 80, pp. 3812-3815
  • Yang, Y.C., Spalholz, B.A., Rabson, M.S., Howley, P.M., Dissociation of transforming and trans-activation functions for bovine papillomavirus type 1 (1985) Nature, 318, pp. 575-577
  • Soeda, E., Ferran, M.C., Baker, C.C., McBride, A.A., Repression of HPV16 early region transcription by the E2 protein (2006) Virology, , in press
  • Mantovani, F., Banks, L., The human papillomavirus E6 protein and its contribution to malignant progression (2001) Oncogene, 20, pp. 7874-7887
  • Munger, K., Phelps, W.C., Bubb, V., Howley, P.M., Schlegel, R., The E6 and E7 genes of the human papillomavirus type 16 together are necessary and sufficient for transformation of primary human keratinocytes (1989) J. Virol, 63, pp. 4417-4421
  • Syrjänen, S., (2005) HPV Today, pp. 8-10. , BYPASS Ediciones, Madrid
  • Simonson, S.J., Difilippantonio, M.J., Lambert, P.F., Two distinct activities contribute to human papillomavirus 16 E6's oncogenic potential (2005) Cancer Res, 65, pp. 8266-8273
  • Kiyono, T., Foster, S.A., Koop, J.I., McDougall, J.K., Galloway, D.A., Klingelhutz, A.J., Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells (1998) Nature, 396, pp. 84-88
  • Barbosa, M.S., Lowy, D.R., Schiller, J.T., Papillomavirus polypeptides E6 and E7 are zinc-binding proteins (1989) J. Virol, 63, pp. 1404-1407
  • Huibregtse, J.M., Scheffner, M., Beaudenon, S., Howley, P.M., A family of proteins structurally and functionally related to the E6-AP ubiquitin-protein ligase (1995) Proc. Natl. Acad. Sci. U.S.A, 92, p. 5249
  • Nomine, Y., Masson, M., Charbonnier, S., Zanier, K., Ristriani, T., Deryckere, F., Sibler, A.P., Trave, G., Structural and functional analysis of E6 oncoprotein: Insights inthe molecular pathways of human papillomavirus-mediated pathogenesis (2006) Mol. Cell, 21, pp. 665-678
  • Daniels, P.R., Sanders, C.M., Maitland, N.J., Characterization of the interactions of human papillomavirus type 16 E6 with p53 and E6-associated protein in insect and human cells (1998) J. Gen. Virol, 79 (PART 3), pp. 489-499
  • Liang, X.H., Volkmann, M., Klein, R., Herman, B., Lockett, S.J., Co-localization of the tumor-suppressor protein p53 and human papillomavirus E6 protein in human cervical carcinoma cell lines (1993) Oncogene, 8, pp. 2645-2652
  • Kanda, T., Watanabe, S., Zanma, S., Sato, H., Furuno, A., Yoshiike, K., Human papillomavirus type 16 E6 proteins with glycine substitution for cysteine in the metal-binding motif (1991) Virology, 185, pp. 536-543
  • Masson, M., Hindelang, C., Sibler, A.P., Schwalbach, G., Trave, G., Weiss, E., Preferential nuclear localization of the human papillomavirus type 16 E6 oncoprotein in cervical carcinoma cells (2003) J. Gen. Virol, 84, pp. 2099-2104
  • Guccione, E., Massimi, P., Bernat, A., Banks, L., Comparative analysis of the intracellular location of the high- and low-risk human papillomavirus oncoproteins (2002) Virology, 293, pp. 20-25
  • Alonso, L.G., Garcia-Alai, M.M., Smal, C., Centeno, J.M., Iacono, R., Castano, E., Gualfetti, P., de Prat-Gay, G., The HPV16 E7 viral oncoprotein self-assembles into defined spherical oligomers (2004) Biochemistry, 43, pp. 3310-3317
  • Schwarz, E., Freese, U.K., Gissmann, L., Mayer, W., Roggenbuck, B., Stremlau, A., zur Hausen, H., Structure and transcription of human papillomavirus sequences in cervical carcinoma cells (1985) Nature, 314, pp. 111-114
  • Uversky, V.N., Natively unfolded proteins: A point where biology waits for physics (2002) Protein Sci, 11, pp. 739-756
  • Nomine, Y., Ristriani, T., Laurent, C., Lefevre, J.F., Weiss, E., Trave, G., Formation of soluble inclusion bodies by hpv e6 oncoprotein fused to maltose-binding protein (2001) Protein Expression Purif, 23, pp. 22-32
  • Nomine, Y., Ristriani, T., Laurent, C., Lefevre, J.F., Weiss, E., Trave, G., A strategy for optimizing the monodispersity of fusion proteins: Application to purification of recombinant HPV E6 oncoprotein (2001) Protein Eng, 14, pp. 297-305
  • Cole, S.T., Danos, O., Nucleotide sequence and comparative analysis of the human papillomavirus type 18 genome. Phylogeny of papillomaviruses and repeated structure of the E6 and E7 gene products (1987) J. Mol. Biol, 193, pp. 599-608. , 28
  • Degenkolbe, R., Gilligan, P., Gupta, S., Bernard, H.U., Chelating agents stabilize the monomeric state of the zinc binding human papillomavirus 16 E6 oncoprotein (2003) Biochemistry, 42, pp. 3868-3873
  • Dippold, W.G., Jay, G., DeLeo, A.B., Khoury, G., Old, L.J., p53 transformation-related protein: Detection by monoclonal antibody in mouse and human cells (1981) Proc. Natl. Acad. Sci. U.S.A, 78, pp. 1695-1699
  • Rotter, V., Abutbul, H., Wolf, D., The presence of p53 transformation-related protein in Ab-MuLV transformed cells is required for their development into lethal tumors in mice (1983) Int. J. Cancer, 31, pp. 315-320
  • Thomas, M., Massimi, P., Navarro, C., Borg, J.P., Banks, L., The hScrib/Dlg apico-basal control complex is differentially targeted by HPV-16 and HPV-18 E6 proteins (2005) Oncogene, 24, pp. 6222-6230
  • Woods, D.F., Wu, J.W., Bryant, P.J., Localization of proteins to the apico-lateral junctions of Drosophila epithelia (1997) Dev. Genet, 20, pp. 111-118

Citas:

---------- APA ----------
García-Alai, M.M., Dantur, K.I., Smal, C., Pietrasanta, L. & De Prat-Gay, G. (2007) . High-risk HPV E6 oncoproteins assemble into large oligomers that allow localization of endogenous species in prototypic HPV-transformed cell lines. Biochemistry, 46(2), 341-349.
http://dx.doi.org/10.1021/bi602457q
---------- CHICAGO ----------
García-Alai, M.M., Dantur, K.I., Smal, C., Pietrasanta, L., De Prat-Gay, G. "High-risk HPV E6 oncoproteins assemble into large oligomers that allow localization of endogenous species in prototypic HPV-transformed cell lines" . Biochemistry 46, no. 2 (2007) : 341-349.
http://dx.doi.org/10.1021/bi602457q
---------- MLA ----------
García-Alai, M.M., Dantur, K.I., Smal, C., Pietrasanta, L., De Prat-Gay, G. "High-risk HPV E6 oncoproteins assemble into large oligomers that allow localization of endogenous species in prototypic HPV-transformed cell lines" . Biochemistry, vol. 46, no. 2, 2007, pp. 341-349.
http://dx.doi.org/10.1021/bi602457q
---------- VANCOUVER ----------
García-Alai, M.M., Dantur, K.I., Smal, C., Pietrasanta, L., De Prat-Gay, G. High-risk HPV E6 oncoproteins assemble into large oligomers that allow localization of endogenous species in prototypic HPV-transformed cell lines. Biochemistry. 2007;46(2):341-349.
http://dx.doi.org/10.1021/bi602457q