Artículo

Alonso, L.G.; García-Alai, M.M.; Smal, C.; Centeno, J.M.; Iacono, R.; Castaño, E.; Gualfetti, P.; De Prat-Gay, G. "The HPV16 E7 Viral Oncoprotein Self-Assembles into Defined Spherical Oligomers" (2004) Biochemistry. 43(12):3310-3317
La versión final de este artículo es de uso interno. El editor solo permite incluir en el repositorio el artículo en su versión post-print. Por favor, si usted la posee enviela a
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

Despite the fact that E7 is a major transforming oncoprotein in papillomavirus, its structure and precise molecular mechanism of action remain puzzling to date. E7 proteins share sequence homology and proteasome targeting properties of tumor suppressors with adenovirus E1A and SV40 T antigen, two other paradigmatic oncoproteins from DNA tumor viruses. High-risk HPV16 E7, a nonglobular dimer with some properties of intrinsically disordered proteins, is capable of undergoing pH-dependent conformational transitions that expose hydrophobic surfaces to the solvent. We found that treatment with a chelating agent produced a protein that can readily assemble into homogeneous spherical particles with an average molecular mass of 790 kDa and a diameter of 50 nm, as determined from dynamic light scattering and electron microscopy. The protein undergoes a substantial conformational transition from coil to β-sheet structure, with concomitant consolidation of tertiary structure as judged by circular dichroism and fluorescence. The assembly process is very slow, in agreement with a substantial energy barrier caused by structural rearrangements. The resulting particles are highly stable, cooperatively folded, and capable of binding both Congo Red and thioflavin T, reporters of repetitive β-sheet structures similar to those found in amyloids, although no fibrillar or insoluble material was observed under our experimental conditions.

Registro:

Documento: Artículo
Título:The HPV16 E7 Viral Oncoprotein Self-Assembles into Defined Spherical Oligomers
Autor:Alonso, L.G.; García-Alai, M.M.; Smal, C.; Centeno, J.M.; Iacono, R.; Castaño, E.; Gualfetti, P.; De Prat-Gay, G.
Filiación:Instituto Leloir, Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Patricias Argentinas 435, (1405) Buenos Aires, Argentina
Departamento de Quimica Biologica, Facultad de Farmacia y Bioquimica, Universidad de Buenos Aires, Junin 956, C1113AAD, Buenos Aires, Argentina
Genencor International, Inc., 925 Page Mill Road, Palo Alto, CA 94304, United States
Palabras clave:Chelation; DNA; Hydrophobicity; Oligomers; pH; Proteins; Self assembly; Tumors; Viruses; Oncoproteins; Spherical materials; Biochemistry; congo red; oligomer; oncoprotein; protein E7; thioflavine; virus protein; virus T antigen; article; beta sheet; cancer inhibition; circular dichroism; conformational transition; DNA tumor virus; electron microscopy; fluorescence; hydrophobicity; light scattering; molecular mechanics; molecular weight; nonhuman; pH measurement; priority journal; protein assembly; protein binding; protein conformation; protein folding; protein stability; protein structure; protein tertiary structure; sequence homology; Simian virus 40; Wart virus; Casein Kinase II; Circular Dichroism; Congo Red; Dimerization; Fluorescent Dyes; Humans; Molecular Weight; Oncogene Proteins, Viral; Papillomaviridae; Phosphorylation; Protein Binding; Protein Structure, Secondary; Protein-Serine-Threonine Kinases; Solubility; Thiazoles; Virus Assembly; Zinc; Adenoviridae; DNA viruses; Human papillomavirus types; Papillomavirus; Simiae; Simian virus 40
Año:2004
Volumen:43
Número:12
Página de inicio:3310
Página de fin:3317
DOI: http://dx.doi.org/10.1021/bi036037o
Título revista:Biochemistry
Título revista abreviado:Biochemistry
ISSN:00062960
CODEN:BICHA
CAS:congo red, 573-58-0, 80701-77-5; thioflavine, 2390-54-7; Casein Kinase II, EC 2.7.1.37; Congo Red, 573-58-0; Fluorescent Dyes; oncogene protein E7, Human papillomavirus type 16; Oncogene Proteins, Viral; Protein-Serine-Threonine Kinases, EC 2.7.1.37; Thiazoles; thioflavin T, 2390-54-7; Zinc, 7440-66-6
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v43_n12_p3310_Alonso

Referencias:

  • Zur Hausen, H., Papillomavirus infections: A major cause of human cancers (1996) Biochim. Biophys. Acta, 1288, pp. F55-F78
  • Yang, L., Mohr, I., Fouts, E., Lim, D.A., Nohaile, M., Botchan, M., The E1 protein of bovine papilloma virus 1 is an ATP-dependent DNA helicase (1993) Proc. Natl. Acad. Sci. U.S.A., 90, pp. 5086-5090
  • Munger, K., The role of human papillomaviruses in human cancers (2002) Front. Biosci., 7, pp. d641-d649
  • Tommasino, M., Crawford, L., Human papillomavirus E6 and E7: Proteins which deregulate the cell cycle (1995) BioEssays, 17, pp. 509-518
  • Phelps, W.C., Yee, C.L., Munger, K., Howley, P.M., The human papillomavirus type 16 E7 gene encodes transactivation and transformation functions similar to those of adenovirus E1A (1988) Cell, 53, pp. 539-547
  • Munger, K., Phelps, W.C., The human papillomavirus E7 protein as a transforming and transactivating factor (1993) Biochim. Biophys. Acta, 1155, pp. 111-123
  • Sato, H., Furuno, A., Yoshiike, K., Expression of human papillomavirus type 16 E7 gene induces DNA synthesis of rat 3Y1 cells (1989) Virology, 168, pp. 195-199
  • Barbosa, M.S., Edmonds, C., Fisher, C., Schiller, J.T., Lowy, D.R., Vousden, K.H., The region of the HPV E7 oncoprotein homologous to adenovirus E1a and Sv40 large T antigen contains separate domains for Rb binding and casein kinase II phosphorylation (1990) EMBO J., 9, pp. 153-160
  • Dyson, N., Howley, P.M., Munger, K., Harlow, E., The human papilloma virus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product (1989) Science, 243, pp. 934-937
  • Zerfass, K., Levy, L.M., Cremonesi, C., Ciccolini, F., Jansen-Durr, P., Crawford, L., Ralston, R., Tommasino, M., Cell cycle-dependent disruption of E2F-p107 complexes by human papillomavirus type 16 E7 (1995) J. Gen. Virol., 76, pp. 1815-1820
  • Chellappan, S., Kraus, V.B., Kroger, B., Munger, K., Howley, P.M., Phelps, W.C., Nevins, J.R., Adenovirus E1A, simian virus 40 tumor antigen, and human papillomavirus E7 protein share the capacity to disrupt the interaction between transcription factor E2F and the retinoblastoma gene product (1992) Proc. Natl. Acad. Sci. U.S.A., 89, pp. 4549-4553
  • 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
  • McIntyre, M.C., Frattini, M.G., Grossman, S.R., Laimins, L.A., Human papillomavirus type 18 E7 protein requires intact Cys-X-X-Cys motifs for zinc binding, dimerization, and transformation but not for Rb binding (1993) J. Virol., 67, pp. 3142-3150
  • Smotkin, D., Wettstein, F.O., The major human papillomavirus protein in cervical cancers is a cytoplasmic phosphoprotein (1987) J. Virol., 61, pp. 1686-1689
  • Munger, K., Basile, J.R., Duensing, S., Eichten, A., Gonzalez, S.L., Grace, M., Zacny, V.L., Biological activities and molecular targets of the human papillomavirus E7 oncoprotein (2001) Oncogene, 20, pp. 7888-7898
  • Dyson, H.J., Wright, P.E., Coupling of folding and binding for unstructured proteins (2002) Curr. Opin. Struct. Biol., 12, pp. 54-60
  • Uversky, V.N., Gillespie, J.R., Fink, A.L., Why are "natively unfolded" proteins unstructured under physiologic conditions? (2000) Proteins, 41, pp. 415-427
  • Alonso, L.G., Garcia-Alai, M.M., Nadra, A.D., Lapena, A.N., Almeida, F.L., Gualfetti, P., Prat-Gay, G.D., High-risk (HPV16) human papillomavirus E7 oncoprotein is highly stable and extended, with conformational transitions that could explain its multiple cellular binding partners (2002) Biochemistry, 41, pp. 10510-10518
  • Mok, Y.K., De Prat Gay, G., Butler, P.J., Bycroft, M., Equilibrium dissociation and unfolding of the dimeric human papillomavirus strain-16 E2 DNA-binding domain (1996) Protein Sci., 5, pp. 310-319
  • Krebs, M.R., Wilkins, D.K., Chung, E.W., Pitkeathly, M.C., Chamberlain, A.K., Zurdo, J., Robinson, C.V., Dobson, C.M., Formation and seeding of amyloid fibrils from wild-type hen lysozyme and a peptide fragment from the β-domain (2000) J. Mol. Biol., 300, pp. 541-549
  • Hunt, J.B., Neece, S.H., Ginsburg, A., The use of 4-(2-pyridylazo)resorcinol in studies of zinc release from Escherichia coli aspartate transcarbamoylase (1985) Anal. Biochem., 146, pp. 150-157
  • Clements, A., Johnston, K., Mazzarelli, J.M., Ricciardi, R.P., Marmorstein, R., Oligomerization properties of the viral oncoproteins adenovirus E1A and human papillomavirus E7 and their complexes with the retinoblastoma protein (2000) Biochemistry, 39, pp. 16033-16045
  • Patrick, D.R., Zhang, K., Defeo-Jones, D., Vuocolo, G.R., Maigetter, R.Z., Sardana, M.K., Oliff, A., Heimbrook, D.C., Characterization of functional HPV-16 E7 protein produced in Escherichia coli (1992) J. Biol. Chem., 267, pp. 6910-6915
  • Pahel, G., Aulabaugh, A., Short, S.A., Barnes, J.A., Painter, G.R., Ray, P., Phelps, W.C., Structural and functional characterization of the HPV16 E7 protein expressed in bacteria (1993) J. Biol. Chem., 268, pp. 26018-26025
  • Clemens, K.E., Brent, R., Gyuris, J., Munger, K., Dimerization of the human papillomavirus E7 oncoprotein in vivo (1995) Virology, 214, pp. 289-293
  • Uversky, V.N., What does it mean to be natively unfolded? (2002) Eur. J. Biochem., 269, pp. 2-12
  • LeVine III, H., Quantification of β-sheet amyloid fibril structures with thioflavin T (1999) Methods Enzymol., 309, pp. 274-284
  • Klunk, W.E., Jacob, R.F., Mason, R.P., Quantifying amyloid by congo red spectral shift assay (1999) Methods Enzymol., 309, pp. 285-305
  • Khurana, R., Uversky, V.N., Nielsen, L., Fink, A.L., Is Congo red an amyloid-specific dye? (2001) J. Biol. Chem., 276, pp. 22715-22721
  • Firzlaff, J.M., Luscher, B., Eisenman, R.N., Negative charge at the casein kinase II phosphorylation site is important for transformation but not for Rb protein binding by the E7 protein of human papillomavirus type 16 (1991) Proc. Natl. Acad. Sci. U.S.A., 88, pp. 5187-5191
  • Zwerschke, W., Jansen-Durr, P., Cell transformation by the E7 oncoprotein of human papillomavirus type 16: Interactions with nuclear and cytoplasmic target proteins (2000) Adv. Cancer Res., 78, pp. 1-29
  • Caspar, D.L., Klug, A., Physical principles in the construction of regular viruses (1962) Cold Spring Harbor Symp. Quant. Biol., 27, pp. 1-24
  • Blundell, T.L., Srinivasan, N., Symmetry, stability, and dynamics of multidomain and multicomponent protein systems (1996) Proc. Natl. Acad. Sci. U.S.A., 93, pp. 14243-14248
  • Kentsis, A., Gordon, R.E., Borden, K.L., Control of biochemical reactions through supramolecular RING domain self-assembly (2002) Proc. Natl. Acad. Sci. U.S.A., 99, pp. 15404-15409
  • Kentsis, A., Gordon, R.E., Borden, K.L., Self-assembly properties of a model RING domain (2002) Proc. Natl. Acad. Sci. U.S.A., 99, pp. 667-672
  • Melnick, A., Licht, J.D., Deconstructing a disease: RARalpha, its fusion partners, and their roles in the pathogenesis of acute promyelocytic leukemia (1999) Blood, 93, pp. 3167-3215
  • 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
  • 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
  • Cole, S.T., Danos, O., Nucleotide sequence and comparative analysis of the human papillomavirus type 18 genome (1987) J. Mol. Biol., 193, pp. 599-608
  • Outten, C.E., O'Halloran, T.V., Femtomolar sensitivity of metalloregulatory proteins controlling zinc homeostasis (2001) Science, 292, pp. 2488-2492

Citas:

---------- APA ----------
Alonso, L.G., García-Alai, M.M., Smal, C., Centeno, J.M., Iacono, R., Castaño, E., Gualfetti, P.,..., De Prat-Gay, G. (2004) . The HPV16 E7 Viral Oncoprotein Self-Assembles into Defined Spherical Oligomers. Biochemistry, 43(12), 3310-3317.
http://dx.doi.org/10.1021/bi036037o
---------- CHICAGO ----------
Alonso, L.G., García-Alai, M.M., Smal, C., Centeno, J.M., Iacono, R., Castaño, E., et al. "The HPV16 E7 Viral Oncoprotein Self-Assembles into Defined Spherical Oligomers" . Biochemistry 43, no. 12 (2004) : 3310-3317.
http://dx.doi.org/10.1021/bi036037o
---------- MLA ----------
Alonso, L.G., García-Alai, M.M., Smal, C., Centeno, J.M., Iacono, R., Castaño, E., et al. "The HPV16 E7 Viral Oncoprotein Self-Assembles into Defined Spherical Oligomers" . Biochemistry, vol. 43, no. 12, 2004, pp. 3310-3317.
http://dx.doi.org/10.1021/bi036037o
---------- VANCOUVER ----------
Alonso, L.G., García-Alai, M.M., Smal, C., Centeno, J.M., Iacono, R., Castaño, E., et al. The HPV16 E7 Viral Oncoprotein Self-Assembles into Defined Spherical Oligomers. Biochemistry. 2004;43(12):3310-3317.
http://dx.doi.org/10.1021/bi036037o