Cytomegalovirus Replicon-Based Regulation of Gene Expression In Vitro and In Vivo

There is increasing evidence for a connection between DNA replication and the expression of adjacent genes. Therefore, this study addressed the question of whether a herpesvirus origin of replication can be used to activate or increase the expression of adjacent genes. Cell lines carrying an episomal vector, in which reporter genes are linked to the murine cytomegalovirus (MCMV) origin of lytic replication (oriLyt), were constructed. Reporter gene expression was silenced by a histone-deacetylase-dependent mechanism, but was resolved upon lytic infection with MCMV. Replication of the episome was observed subsequent to infection, leading to the induction of gene expression by more than 1000-fold. oriLyt-based regulation thus provided a unique opportunity for virus-induced conditional gene expression without the need for an additional induction mechanism. This principle was exploited to show effective late trans-complementation of the toxic viral protein M50 and the glycoprotein gO of MCMV. Moreover, the application of this principle for intracellular immunization against herpesvirus infection was demonstrated. The results of the present study show that viral infection specifically activated the expression of a dominant-negative transgene, which inhibited viral growth. This conditional system was operative in explant cultures of transgenic mice, but not in vivo. Several applications are discussed

Soybean Trihelix Transcription Factors GmGT-2A and GmGT-2B Improve Plant Tolerance to Abiotic Stresses in Transgenic Arabidopsis

BACKGROUND:Trihelix transcription factors play important roles in light-regulated responses and other developmental processes. However, their functions in abiotic stress response are largely unclear. In this study, we identified two trihelix transcription factor genes GmGT-2A and GmGT-2B from soybean and further characterized their roles in abiotic stress tolerance. FINDINGS:Both genes can be induced by various abiotic stresses, and the encoded proteins were localized in nuclear region. In yeast assay, GmGT-2B but not GmGT-2A exhibits ability of transcriptional activation and dimerization. The N-terminal peptide of 153 residues in GmGT-2B was the minimal activation domain and the middle region between the two trihelices mediated the dimerization of the GmGT-2B. Transactivation activity of the GmGT-2B was also confirmed in plant cells. DNA binding analysis using yeast one-hybrid assay revealed that GmGT-2A could bind to GT-1bx, GT-2bx, mGT-2bx-2 and D1 whereas GmGT-2B could bind to the latter three elements. Overexpression of the GmGT-2A and GmGT-2B improved plant tolerance to salt, freezing and drought stress in transgenic Arabidopsis plants. Moreover, GmGT-2B-transgenic plants had more green seedlings compared to Col-0 under ABA treatment. Many stress-responsive genes were altered in GmGT-2A- and GmGT-2B-transgenic plants. CONCLUSION:These results indicate that GmGT-2A and GmGT-2B confer stress tolerance through regulation of a common set of genes and specific sets of genes. GmGT-2B also affects ABA sensitivity

Beam Energy Dependence of Jet-Quenching Effects in Au plus Au Collisions at root s(NN)=7.7, 11.5, 14.5, 19.6, 27, 39, and 62.4 GeV

We report measurements of the nuclear modification factor, $R_{ \mathrm{CP}}$, for charged hadrons as well as identified $\pi^{+(-)}$, $K^{+(-)}$, and $p(\overline{p})$ for Au+Au collision energies of $\sqrt{s_{_{ \mathrm{NN}}}}$ = 7.7, 11.5, 14.5, 19.6, 27, 39, and 62.4 GeV. We observe a clear high-$p_{\mathrm{T}}$ net suppression in central collisions at 62.4 GeV for charged hadrons which evolves smoothly to a large net enhancement at lower energies. This trend is driven by the evolution of the pion spectra, but is also very similar for the kaon spectra. While the magnitude of the proton $R_{ \mathrm{CP}}$ at high $p_{\mathrm{T}}$ does depend on collision energy, neither the proton nor the anti-proton $R_{ \mathrm{CP}}$ at high $p_{\mathrm{T}}$ exhibit net suppression at any energy. A study of how the binary collision scaled high-$p_{\mathrm{T}}$ yield evolves with centrality reveals a non-monotonic shape that is consistent with the idea that jet-quenching is increasing faster than the combined phenomena that lead to enhancement.We report measurements of the nuclear modification factor RCP for charged hadrons as well as identified π+(-), K+(-), and p(p¯) for Au+Au collision energies of sNN=7.7, 11.5, 14.5, 19.6, 27, 39, and 62.4 GeV. We observe a clear high-pT net suppression in central collisions at 62.4 GeV for charged hadrons which evolves smoothly to a large net enhancement at lower energies. This trend is driven by the evolution of the pion spectra but is also very similar for the kaon spectra. While the magnitude of the proton RCP at high pT does depend on the collision energy, neither the proton nor the antiproton RCP at high pT exhibit net suppression at any energy. A study of how the binary collision-scaled high-pT yield evolves with centrality reveals a nonmonotonic shape that is consistent with the idea that jet quenching is increasing faster than the combined phenomena that lead to enhancement