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Smal, C.; Wetzler, D.E.; Dantur, K.I.; Chemes, L.B.; Garcia-Alai, M.M.; Dellarole, M.; Alonso, L.G.; Gaston, K.; De Prat-Gay, G. "The human papillomavirus E7-E2 interaction mechanism in vitro reveals a finely tuned system for modulating available E7 and E2 proteins" (2009) Biochemistry. 48(50):11939-11949
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Abstract:

Transcription of the human papillomavirus E7 oncoprotein is negatively controlled by the viral E2 protein, and loss of this repression leads to irreversible transformation and carcinogenesis. Here we show that interaction of the HPV16 E7 protein with the DNA binding domain of the E2 protein (E2C) leads to ionic strength-dependent hetero-oligomerization even at the lowest concentrations measurable. Titration experiments followed by light scattering and native gel electrophoresis show insoluble oligomeric complexes with a ≥2000 nm diameter and intermediate soluble complexes 40 and 115 nm in diameter, respectively, formed in excess of E2C. Adiscrete oligomeric soluble complex formed in excess of E7 displays a diameter of 12 nm. The N-terminal domain of E7 interacts with E2C with a KD of 0.1 μM, where the stretch of residues 25-40 of E7, encompassing both a PEST motif and phosphorylation sites, is sufficient for the interaction. Displacement of the soluble E7-E2C complex by an E2 site DNA duplex and site-directed mutagenesis indicate that the protein-protein interface involves the DNA binding helix of E2. The formation of complexes of different sizes and properties in excess of either of the viral proteins reveals a finely tuned mechanism that could regulate the intracellular levels of both proteins as infection and transformation progress. Sequestering E2 into E7-E2 oligomers provides a possible additional route to uncontrolled E7 expression, in addition and prior to the disruption of the E2 gene during viral integration into the host genome. © 2009 American Chemical Society.

Registro:

Documento: Artículo
Título:The human papillomavirus E7-E2 interaction mechanism in vitro reveals a finely tuned system for modulating available E7 and E2 proteins
Autor:Smal, C.; Wetzler, D.E.; Dantur, K.I.; Chemes, L.B.; Garcia-Alai, M.M.; Dellarole, M.; Alonso, L.G.; Gaston, K.; De Prat-Gay, G.
Filiación:Fundación Instituto Leloir, Instituto de Investigaciones Bioquímicas, CONICET, Patricias Argentinas 435, 1405 Buenos Aires, Argentina
Facultad de Ciencias Exactas Y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom
Medical Research Council, Centre for Protein Engineering, Cambridge CB2 0QH, United Kingdom
Palabras clave:Different sizes; DNA binding; DNA binding domain; DNA duplexes; E7 oncoprotein; Gel electrophoresis; Hetero-oligomerization; Human papillomavirus; In-vitro; Interaction mechanisms; Intracellular levels; N-terminal domains; Oligomeric complexes; Phosphorylation sites; Protein-protein interface; Site directed mutagenesis; Soluble complexes; Viral proteins; Binding sites; Complexation; DNA; Electrophoresis; Enzyme activity; Genes; Ionic strength; Nucleic acids; Oligomerization; Oligomers; Phosphorylation; Titration; Transcription; Proteins; protein E7; protein VP2; amino terminal sequence; article; DNA binding; gel electrophoresis; human; in vitro study; ionic strength; light scattering; nonhuman; oligomerization; phosphorylation; priority journal; protein interaction; titrimetry; Wart virus; Amino Acid Sequence; Cell Line, Tumor; Cell Proliferation; DNA-Binding Proteins; Human papillomavirus 16; Humans; Molecular Sequence Data; Oncogene Proteins, Viral; Protein Structure, Tertiary; ras Proteins; Virus Integration; Human papillomavirus
Año:2009
Volumen:48
Número:50
Página de inicio:11939
Página de fin:11949
DOI: http://dx.doi.org/10.1021/bi901415k
Título revista:Biochemistry
Título revista abreviado:Biochemistry
ISSN:00062960
CODEN:BICHA
CAS:DNA-Binding Proteins; E2 protein, Human papillomavirus type 16; Oncogene Proteins, Viral; oncogene protein E7, Human papillomavirus type 16; ras Proteins, 3.6.5.2
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v48_n50_p11939_Smal

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Citas:

---------- APA ----------
Smal, C., Wetzler, D.E., Dantur, K.I., Chemes, L.B., Garcia-Alai, M.M., Dellarole, M., Alonso, L.G.,..., De Prat-Gay, G. (2009) . The human papillomavirus E7-E2 interaction mechanism in vitro reveals a finely tuned system for modulating available E7 and E2 proteins. Biochemistry, 48(50), 11939-11949.
http://dx.doi.org/10.1021/bi901415k
---------- CHICAGO ----------
Smal, C., Wetzler, D.E., Dantur, K.I., Chemes, L.B., Garcia-Alai, M.M., Dellarole, M., et al. "The human papillomavirus E7-E2 interaction mechanism in vitro reveals a finely tuned system for modulating available E7 and E2 proteins" . Biochemistry 48, no. 50 (2009) : 11939-11949.
http://dx.doi.org/10.1021/bi901415k
---------- MLA ----------
Smal, C., Wetzler, D.E., Dantur, K.I., Chemes, L.B., Garcia-Alai, M.M., Dellarole, M., et al. "The human papillomavirus E7-E2 interaction mechanism in vitro reveals a finely tuned system for modulating available E7 and E2 proteins" . Biochemistry, vol. 48, no. 50, 2009, pp. 11939-11949.
http://dx.doi.org/10.1021/bi901415k
---------- VANCOUVER ----------
Smal, C., Wetzler, D.E., Dantur, K.I., Chemes, L.B., Garcia-Alai, M.M., Dellarole, M., et al. The human papillomavirus E7-E2 interaction mechanism in vitro reveals a finely tuned system for modulating available E7 and E2 proteins. Biochemistry. 2009;48(50):11939-11949.
http://dx.doi.org/10.1021/bi901415k