Cooperative Activation of Human Papillomavirus Type 8 Gene Expression by the E2 Protein and the Cellular Coactivator p300

The E2 proteins of papillomaviruses (PV) bind to the coactivator CBP/p300 as do many other transcription factors, but the precise role of CBP/p300 in E2-specific functions is not yet understood. We show that the E2 protein of human PV type 8 (HPV8) directly binds to p300. Activation of HPV8 gene expression by low amounts of HPV8 E2 was stimulated up to sevenfold by coexpression of p300. The interaction between E2 and p300 may play a role in differentiation-dependent activation of PV gene expression, since we can show that the expression level of p300 increases during keratinocyte differentiation. Surprisingly, sequence-specific binding of E2 to its recognition sites within the regulatory region of HPV8 is not necessary for this cooperation, indicating that E2 can be recruited to the promoter via protein-protein interaction. HPV8 E2 binds via its N-terminal activation domain (AD), its C-terminal DNA binding domain (DBD), and its internal hinge region to p300 in vitro. Transient-transfection assays revealed that the AD is necessary and sufficient for cooperative activation with p300. However, we provide evidence that the interaction of the hinge and the DBD of HPV8 E2 with p300 may contribute. Our data suggest an important role of p300 in regulation of HPV8 gene expression and reveal a new mechanism by which E2 may be recruited to a promoter to activate transcription without sequence specific DNA binding

Broadband analysis and self-control of spectral fluctuations in a passively phase-stable Er-doped fiber frequency comb

Carrier-envelope and optical phase noise of a femtosecond frequency comb based on Er-doped fiber technology is investigated and minimized without exploiting active external references. Ultrabroadband, coherent, and tailorable supercontinua are generated in a highly nonlinear germanosilicate fiber assembly. Difference frequency mixing between comb modes in their spectral extrema passively eliminates the carrier-envelope phase slip. This step generates an inherently offset-free comb with a relative frequency stability better than 10−21. In contrast, the phase fluctuations at the carrier frequency of 193 THz are increased as compared to the fundamental comb. Their level matches the value found by parabolic extrapolation of the phase noise of the fundamental comb to zero frequency. The latter is unambiguously accessible by means of any beat note centered at the carrier-envelope offset frequency. All these findings rely on strong correlations between the comb modes that are quantitatively described by an elastic tape model, underlining the deterministic character of the processes involved. The superior optical phase noise of the fundamental comb is transferred to the difference-frequency comb while not compromising the inherent cancellation of the carrier-envelope offset frequency. In this way, the optical linewidth of the passively phase-locked comb is reduced from 100 kHz to a measured value of 5 kHz, which is limited by the cw laser reference used for out-of-loop characterization.publishe