Structural and thermodynamic basis for the enhanced transcriptional control by the human papillomavirus strain-16 E2 protein.
Strain 16 of the human papillomavirus is responsible for the largest number of cases of cervical cancers linked to this virus, and the E2 protein is the transcriptional regulator of all viral genes. We present the first structure for the DNA binding domain of HPV16 E2 bound to DNA, and in particular, to a natural cognate sequence. The NMR structure of the protein backbone reveals that the overall conformation remains virtually unchanged, and chemical shift analysis of the protein bound to a shorter DNA duplex uncovered a contact out of the minimal E2 DNA binding site, made by lysine 349. This contact was confirmed by titration calorimetry and mutagenesis, with a contribution of 1.0 kcal mol(-)(1) to binding energy. HPV16 E2 has the highest DNA binding affinity and exerts a strict transcriptional control, translated into the repression of the E6 and E7 oncogenes. These novel features provide the structural and thermodynamic basis for this tight transcriptional control, the loss of which correlates with carcinogenesis.
Cicero
Daniel O
DO
Instituto Leloir and CONICET, University of Rome Tor Vergata, via della Ricerca Scientifica, 00133 Rome, Italy Tor Vergata.
Nadra
Alejandro D
AD
Eliseo
Tommaso
T
Dellarole
Mariano
M
Paci
Maurizio
M
de Prat-Gay
Gonzalo
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009841:Oligonucleotides; D009856:Oncogene Proteins, Viral / Q000737:chemistry* / Q000502:physiology; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013816:Thermodynamics; D014158:Transcription, Genetic / Q000502:physiology*
D004268:DNA-Binding Proteins; C054855:E2 protein, Human papillomavirus type 16; D009841:Oligonucleotides; D009856:Oncogene Proteins, Viral