CAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene-5

<p><b>Copyright information:</b></p><p>Taken from "cAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene"</p><p>BMC Molecular Biology 2005;6():2-2.</p><p>Published online 19 Jan 2005</p><p>PMCID:PMC548273.</p><p></p>nal activation domain of CREB2 and the bZIP domain of ATF2, responsible for dimerization and DNA-binding. (B, C, D) HepG2 cells were transfected with one of the reporter plasmids pG6PCRE1/CRE2luc, pG6PCRE1mut/CRE2luc pG6PCRE1/CRE2mutluc (B), pG6PCRE1luc, pG6PCRE2luc (C), or pTNFα(CRE/AP1)luc (D), the pRSVβ internal reference plasmid, and either the "empty" expression vector pCMV5 or an expression vector encoding C2/ATF2 (100 ng expression plasmid/plate). Lysates were prepared forty-eight hours post-transfection and β-galactosidase and luciferase activities were measured. The mean +/- SD is depicted

CAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene-3

<p><b>Copyright information:</b></p><p>Taken from "cAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene"</p><p>BMC Molecular Biology 2005;6():2-2.</p><p>Published online 19 Jan 2005</p><p>PMCID:PMC548273.</p><p></p>e wild-type enzyme is myristylated as indicated (Myr). The sequence of the nuclear localization signal derived from the SV40 large T antigen and the triple FLAG epitope in NLSCα are shown. (B) Western Blot analysis of HepG2 cells transfected with an expression vector encoding NLSCα. As a control, extracts from mock transfected HepG2 cells were analyzed. Western blots were probed with an antibody against the FLAG-tag. (C) Modular structure of the GAL4-CREB fusion protein. The protein consists of the DNA-binding domain of GAL4 (amino acids 1–147) and the activation domain of CREB (amino acids 1–281). The reporter plasmid pUASluc contains a transcription unit encompassing the luciferase open reading frame and a minimal promoter that consists of five copies of the upstream activating sequence (UAS), a TATA box derived from the HIV long terminal repeat and the initiator element from the adenovirus major late promoter. The reporter plasmid pUASluc (1 μg/plate) was transfected into HepG2 cells together with the pRSVβ internal standard plasmid (2 μg/plate) and either an expression vector encoding the DNA binding domain of GAL4 (plasmid pM1) or the GAL4-CREB fusion protein (1 μg plasmid/plate). In addition, cells were transfected with an expression vector encoding either the wild-type (Cα) or mutated (NLSCα) form of cAMP-dependent protein kinase (100 ng/plate). Forty-eight hours post-transfection cell extracts were prepared and the β-galactosidase and luciferase activities of these extracts were determined

CAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene-4

<p><b>Copyright information:</b></p><p>Taken from "cAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene"</p><p>BMC Molecular Biology 2005;6():2-2.</p><p>Published online 19 Jan 2005</p><p>PMCID:PMC548273.</p><p></p> (C) (1 μg/plate) was transfected into HepG2 cells together with the pRSVβ internal standard plasmid (2 μg/plate) and the "empty" expression vector pCMV5 or an expression vector encoding either CREB, CREBS133A, or K-CREB (20 ng plasmid/plate). In addition, an expression vector encoding NLSCα (100 ng/plate) was transfected. Forty-eight hours post-transfection cell extracts were prepared and the β-galactosidase and luciferase activities of these extracts were determined. The data are presented as the ratio of luciferase activity (light units) to β-galactosidase units (OD units) measured in the cell extracts. The mean +/- SD is depicted

CAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene-1

<p><b>Copyright information:</b></p><p>Taken from "cAMP response element binding protein (CREB) activates transcription via two distinct genetic elements of the human glucose-6-phosphatase gene"</p><p>BMC Molecular Biology 2005;6():2-2.</p><p>Published online 19 Jan 2005</p><p>PMCID:PMC548273.</p><p></p>The chimeric bZIP protein C2/CREB consists of the constitutively active transcriptional activation domain of CREB2 and the bZIP domain of CREB, responsible for dimerization and DNA-binding. (B) Western Blot analysis of HepG2 cells transfected with an expression vector encoding C2/CREB. As a control, extracts from mock transfected HepG2 cells were analyzed. Western blots were probed with an antibody against the FLAG-tag