Upstream activating sequence

About: Upstream activating sequence is a research topic. Over the lifetime, 1633 publications have been published within this topic receiving 100112 citations.

Collapsin response mediator protein-1 (CRMP-1) transcriptional regulatory nucleic acid sequences

Abstract: This invention provides a transcription unit which is isolated from the upstream nucleic acid sequence of the collapsing response mediator protein-1 (CRMP-1) gene, an invasion-suppressor gene. The transcription unit contains a nucleic acid regulatory sequence which demonstrates promoter and/or regulatory activities (such as providing a transcription factor binding site) to enhance the expression of the CRMP-1 and/or a reporter protein. The invention also provides a DNA construct containing this transcription unit which can be transfected into a host cell. Additionally, the invention provides methods to enhance the expression of CRMP-1 and/or the reporter protein. The over-expression of CRMP-1 in a cancer cell can inhibit the metastasis of the cancer cell.

Cis and trans-acting regulatory elements required for regulation of the CPS1 gene in Saccharomyces cerevisiae

Abstract: To clarify the transcriptional regulation by nutrient limitation of the gene encoding carboxypepti- dase yscS in Saccharomyces cerevisiae (CPSI), we per- formed an analysis of its 5' nonc0ding region. In dele- tion experiments a sequence located between positions -644 and -591 was found to be responsible for tran- scriptional repression of the CPS1 gene in yeast cells grown on rich nitrogen sources. Furthermore, a 162 bp fragment spanning positions - 644 to - 482 of the pro- moter of the CPS1 gene repressed gene expression when placed 3' t0 the upstream activation sequence (UAS) of the heterologous gene CYC1. A fragment containing this putative upstream repression sequence (URS) was shown specifically to bind protein from a yeast extract as demonstrated by gel retardation experiments. Al- though a sequence mediating the control of gene expres- sion by GCN4 was found within the URS element, the GCN4 gene product is not required for DNA-binding activity. In addition, at least three other upstream acti- vation UASs responsible for the activation of CPS1 ex- pression by glucose under nitrogen starvation condi- tions were found to be located between positions -673 and -644, -482 and -353, and -243 and -186, re- spectively. The putative mechanism of the nitrogen lim- itation-dependent of CPS1 expression via these regulatory elements is discussed.

The presence of a G-quadruplex prone sequence upstream of a minimal promoter increases transcriptional activity in the yeast S. cerevisiae

Abstract: Non-canonical secondary structures in DNA are increasingly being revealed as critical players in DNA metabolism, including modulating the accessibility and activity of promoters. These structures comprise the so-called G-quadruplexes (G4s) that are formed from sequences rich in guanine bases. Using a well-defined transcriptional reporter system, we sought to systematically investigate the impact of the presence of G4 structures on transcription in yeast S. cerevisiae. To this aim, different G4 prone sequences were modeled to vary the chance of intramolecular G4 formation, analyzed in vitro by Thioflavin T binding test and circular dichroism and then placed at the yeast ADE2 locus on chromosome XV, downstream and adjacent to a P53 response element (RE) and upstream from a minimal CYC1 promoter and Luciferase 1 (LUC1) reporter gene in isogenic strains. While the minimal CYC1 promoter provides for basal reporter activity, the P53 RE enables LUC1 transactivation under the control of the human P53 family proteins expressed under the inducible GAL1 promoter. Thus, the impact of the different G4 prone sequences on both basal and P53 family proteins dependent expression was measured after shifting the yeast cells onto galactose containing medium. The results showed that the presence of G4 prone sequences upstream of a yeast minimal promoter can increase its basal activity proportionally to their potential to form intramolecular G4 structures; consequently, this improved accessibility, when present near the target binding site of P53 family transcription factors can be exploited in order to regulate the transcriptional activity of P53, P63 and P73 proteins.

Purification and functional characterization of a cellular transcription factor that binds to an enhancer element within the adenovirus early EIla promoter (affinity chromatography/band-shift/gene regulation/HeLa cells/in vitro transcription)

Abstract: The adenovirus EIa-inducible early EIIa (EIIaE) promoter is comprised of several sequence elements essential for constitutive and Induced expression. We report here the purification of the host-cell factor that interacts with the major upstream element of this promoter, extending between positions -90 and -70 with respect to the main EIIaE cap site and exhibiting enhancer properties. The puri- fied factor, which corresponds to a 40- to 43-kDa polypeptide, specifically binds to its recognition site and stimulates EIIaE promoter activity when added to an in vitro transcription system, reconstituted from purified factors and RNA polymer- ase. The implication of this factor in the control of the other adenovirus early genes is discussed. Efficient transcription of the adenovirus early transcription units requires the presence of the viral pre-early EIa gene products (1, 2). The mechanism of this transactivation of the early transcription unit is still poorly understood. Extensive deletion and linker scanning mutational analysis (3-5) of the EIa-inducible EIIa early (EIIaE) promoter has indicated that EIa responsiveness involves the same promoter sequences as those required for uninduced expression, suggesting that the same host-cell transcription factors are required in each case. DNA binding studies have identified several of these factors, among which those recognizing sequence elements located between -90 and - 70 (EIIAE-EF (6), EIIaE-B (7,