Navegar

Colección

Datos Estadísticas

Artículo

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:

The C-terminal DNA binding domain of the E2 protein is involved in transcriptional regulation and DNA replication in papillomaviruses. At low ionic strength, the domain has a tendency to form aggregates, a process readily reversible by the addition of salt. While fluorescence anisotropy measurements show a 1:1 stoichiometry at pH 5.5, we observed that a second HPV-16 E2 C-terminal dimer can bind per DNA site at pH 7.0. This was confirmed by displacement of bis-ANS binding, tryptophan fluorescence, native electrophoresis, and circular dichroism. The two binding events are nonequivalent, with a high-affinity binding involving one E2C dimer per DNA molecule with a K(D) of 0.18 ± 0.02 nM and a lower affinity binding mode of 2.0 ± 0.2 nM. The bovine (BPV-1) E2 C-terminal domain binds to an HPV-16 E2 site with 350-fold lower affinity than the human cognate domain and binds 7-fold less tightly even to a bovine-derived DNA site. The ability to discriminate between cognate and noncognate sequences is 50-fold higher for the human domain, and the latter is 180-fold better than the bovine at discriminating specific from nonspecific DNA. A substantial conformational change in bound DNA is observed by near-UV circular dichroism. The bovine domain imposes a different DNA conformation than that caused by the human counterpart, which could be explained by a more pronounced bent. Structure-function differences and biochemical properties of the complexes depend on the protein domain rather than on the DNA, in line with crystallographic evidence. Despite the strong sequence homology and overall folding topology, the differences observed may explain the distinctive transcriptional regulation in bovine and human viruses.

Registro:

Documento: Artículo
Título:Distinctive cognate sequence discrimination, bound DNA conformation, and binding modes in the E2 C-terminal domains from prototype human and bovine papillomaviruses
Autor:Ferreiro, D.U.; Lima, L.M.T.R.; Nadra, A.D.; Alonso, L.G.; Goldbaum, F.A.; De Prat-Gay, G.
Filiación:Instituto de Investigaciones Bioquímicas Fundación Campomar, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Patricias Argentinas 435, (1405) Buenos Aires, Argentina
Faculdade de Farmacia, Universidade Federal do Rio de Janeiro, Cidade Universitaria, Rio de Janeiro 21941-590, Brazil
Palabras clave:article; carboxy terminal sequence; conformational transition; dimerization; DNA conformation; DNA replication; ionic strength; nonhuman; oligomerization; priority journal; protein domain; protein folding; sequence analysis; transactivation; transcription regulation; Adenovirus E2 Proteins; Animals; Base Sequence; Bovine papillomavirus 1; Cattle; Circular Dichroism; Consensus Sequence; DNA; Humans; Nucleic Acid Conformation; Osmolar Concentration; Papillomaviridae; Peptide Fragments; Protein Binding; Protein Folding; Protein Structure, Tertiary; Solutions
Año:2000
Volumen:39
Número:47
Página de inicio:14692
Página de fin:14701
DOI: http://dx.doi.org/10.1021/bi001694r
Título revista:Biochemistry
Título revista abreviado:Biochemistry
ISSN:00062960
CODEN:BICHA
CAS:Adenovirus E2 Proteins; DNA, 9007-49-2; Peptide Fragments; Solutions
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v39_n47_p14692_Ferreiro

Referencias:

  • Tan, S., Richmond, T.J., (1998) Curr. Opin. Struct. Biol., 8, pp. 41-48
  • Edwards, A.M., Bochkarev, A., Frappier, L., (1998) Curr. Opin. Struct. Biol., 8, pp. 49-53
  • McBride, A., Romanczuk, H., Howley, P., (1991) J. Biol. Chem., 266, pp. 18411-18414
  • Piirsoo, M., Ustav, E., Mandel, T.A.S., Stenlund, A., Ustav, M., (1996) EMBO J., 15, pp. 1-11
  • Laimins, L.A., (1998) Human Tumor Viruses, , ASM Press, Washington, DC
  • Giri, I., Yaniv, M., (1988) EMBO J., 7, pp. 2823-2829
  • Gauthier, J.M., Dillner, J., Yaniv, M., (1991) Nucleic Acids Res., 19, pp. 7073-7079
  • Skiadopoulos, M.H., McBride, A.A., (1998) J. Virol., 72, pp. 2079-2088
  • Nishimura, A., Ono, T., Ishimoto, A., Dowhanick, J.J., Frizzell, M.A., Howley, P.M., Sakai, H., (2000) J. Virol., 75, pp. 3752-3760
  • Hegde, R., Grossman, S., Laimins, L., Sigler, P., (1992) Nature, 359, pp. 505-512
  • Bochkarev, A., Barwell, J.A., Pfuetzner, R.A., Bochkareva, E., Frappier, L., Edwards, A.M., (1996) Cell, 84, pp. 791-800
  • Linag, H., Petros, A.M., Meadows, R., Yoon, H.S., Egan, D.A., Walter, K., Holzman, T.F., Fesik, S.W., (1996) Biochemistry, 35, pp. 2095-2103
  • Hegde, R.S., Androphy, E.J., (1998) J. Mol. Biol., 284, pp. 1479-1489
  • Hegde, R.S., Wang, A.F., Kim, S.S., Schapira, M., (1998) J. Mol. Biol., 276, pp. 797-808
  • Li, R., Knight, J., Bream, G., Stenlund, A., Botchan, M., (1989) Genes Dev., 3, pp. 510-526
  • Monini, P., Grossman, S.R., Pepinsky, B., Androphy, E.J., Laimins, L.A., (1991) J. Virol., 65, pp. 2124-2130
  • Sanders, C.M., Maitland, N.J., (1994) Nucleic Acids Res., 22, pp. 4890-4897
  • Tan, S., Leong, L.E., Walker, P.A., Bernard, H., (1994) J. Virol., 68, pp. 6411-6420
  • Chao, S.F., Rocque, W.J., Daniel, S., Czyzyk, L.E., Phelps, W.C., Alexander, K.A., (1999) Biochemistry, 38, pp. 4586-4594
  • Hill, J.J., Royer, C.A., (1997) Methods Enzymol., 278, pp. 390-416
  • Mok, Y.K., De Prat Gay, G., Butler, P.J., Bycroft, M., (1996) Protein Sci., 5, pp. 310-319
  • Mok, Y.K., Bycroft, M., De Prat Gay, G., (1996) Nat. Struct. Biol., 3, pp. 711-717
  • Lima, L.M., De Prat Gay, G., (1997) J. Biol. Chem., 272, pp. 19295-19303
  • Clackson, T., Detlef, G., Jones, P., (1991), p. 202. , PCR, a Practical Approach (McPherson, M., Quirke, P., and Taylor, G., Eds.). IRL Press, Oxford; Pace, C.N., Vajdos, F., Lanette, F., Grimsley, G., Gray, T., (1995) Protein Sci., 4, pp. 2411-2423
  • Alexander, K.A., Phelps, W.C., (1996) Biochemistry, 35, pp. 9864-9872
  • Gray, D., (1996) Circular Dichroism and the Conformational Analysis of Biomolecules, , Plenum Press, New York
  • Ausbel, F., Brent, R., Kingston, R.E., Moore, D.D., Siedman, J.G., Smith, J.G., Struhl, K., (1997) Short Protocols in Molecular Biology, , John Wiley and Sons, New York
  • Thain, A., Webster, K., Emery, D., Clarke, A.R., Gaston, K., (1997) J. Biol. Chem., 272, pp. 8236-8242
  • Hines, C.S., Meghoo, C., Shetty, S., Biburger, M., Brenowitz, M., Hegde, R.S., (1998) J. Mol. Biol., 276, pp. 809-818
  • Foguel, D., Silva, J.L., De Prat Gay, G., (1998) J. Biol. Chem., 273, pp. 9050-9057
  • Pepinsky, R.B., Prakash, S.S., Corina, K., Grossel, M.J., Barsoum, J., Androphy, E.J., (1997) J. Virol., 71, pp. 828-831
  • Sevenich, F.W., Langowski, J., Weiss, V., Rippe, K., (1998) Nucleic Acids Res., 26, pp. 1373-1381
  • Dekker, J., Kanellopoulos, P.N., Loonstra, A.K., Van Oosterhout, J.A., Leonard, K., Tucker, P.A., Van Der Vliet, P.C., (1997) EMBO J., 16, pp. 1455-1463
  • Wyman, C., Rombel, I., North, A.K., Bustamante, C., Kustu, S., (1997) Science, 275, pp. 1658-1661
  • Margeat, E., Le Grimellec, C., Royer, C.A., (1998) Biophys. J., 75, pp. 2712-2720
  • Czernik, P.J., Shin, D.S., Hurlburt, B.K., (1994) J. Biol. Chem., 269, pp. 27869-27875
  • Grillo, A.O., Brown, M.P., Royer, C.A., (1999) J. Mol. Biol., 287, pp. 539-554
  • Rozenberg, H., Rabinovich, D., Frolow, F., Hegde, R.S., Shakked, Z., (1998) Proc. Natl. Acad. Sci. U.S.A., 95, pp. 15194-15199
  • Knight, J.D., Li, R., Botchan, M., (1991) Proc. Natl. Acad. Sci. U.S.A., 88, pp. 3204-3208

Citas:

---------- APA ----------
Ferreiro, D.U., Lima, L.M.T.R., Nadra, A.D., Alonso, L.G., Goldbaum, F.A. & De Prat-Gay, G. (2000) . Distinctive cognate sequence discrimination, bound DNA conformation, and binding modes in the E2 C-terminal domains from prototype human and bovine papillomaviruses. Biochemistry, 39(47), 14692-14701.
http://dx.doi.org/10.1021/bi001694r
---------- CHICAGO ----------
Ferreiro, D.U., Lima, L.M.T.R., Nadra, A.D., Alonso, L.G., Goldbaum, F.A., De Prat-Gay, G. "Distinctive cognate sequence discrimination, bound DNA conformation, and binding modes in the E2 C-terminal domains from prototype human and bovine papillomaviruses" . Biochemistry 39, no. 47 (2000) : 14692-14701.
http://dx.doi.org/10.1021/bi001694r
---------- MLA ----------
Ferreiro, D.U., Lima, L.M.T.R., Nadra, A.D., Alonso, L.G., Goldbaum, F.A., De Prat-Gay, G. "Distinctive cognate sequence discrimination, bound DNA conformation, and binding modes in the E2 C-terminal domains from prototype human and bovine papillomaviruses" . Biochemistry, vol. 39, no. 47, 2000, pp. 14692-14701.
http://dx.doi.org/10.1021/bi001694r
---------- VANCOUVER ----------
Ferreiro, D.U., Lima, L.M.T.R., Nadra, A.D., Alonso, L.G., Goldbaum, F.A., De Prat-Gay, G. Distinctive cognate sequence discrimination, bound DNA conformation, and binding modes in the E2 C-terminal domains from prototype human and bovine papillomaviruses. Biochemistry. 2000;39(47):14692-14701.
http://dx.doi.org/10.1021/bi001694r