LÄHDE:
Trim JE, Samra SK,et al. Upstream tissue inhibitor of metalloproteinases-1 (TIMP-1) element-1, a novel and essential regulatory DNA motif in the human TIMP-1 gene promoter, directly interacts with a 30-kDa nuclear protein. J Biol Chem. 2000 Mar 3;275(9):6657-63. UK
Tiivistelmä, Abstract
- TIMP-1 proteiinin ja sen mRNA :n ylössäätymää tavataan ihmisen taudeissa, kuten syövissä ja kudosfibrooseissa .
- . TIMP-1 geenin ilmentymä välittyy pääasiassa geenitranskription tasolta ja siihen osallistuu usea hyvin tunnettu transkriptiotekijä kuten ne , jotka kuuluvat AP-1-, STAT-, Pea3/Ets perheisiin.
- Tässä tutkimuksessa käytetty DNAaasi-1 identifioi yhden uuden säätelyelementin(5'-TGTGGTTTCCG-3') ihmisen TIMP-1 geeni promoottorista nimeltään upstream TIMP-1 element-1 (UTE-1). Havaittiin, että se on essentielli ihmisen TIMP-1 promoottorin transkriptioaktiivisuudelle. UTE- 1 tekee suoran interaktion 30 kDa tumaproteiinin kanssa jota esiintyi kaikissa testatuissa soluissa. Tutkijat ovat sitä mieltä että tämä UTE-1 on uusi säätelyelementti, joka yhdessä kombinoituneena sen soluperäiseen sitovaan proteiiniinsa voi olla tärkeä komponentti TIMP-1 ilmenemän kontrollimekanismissa normaaleissa ja patologisissa tiloissa.
Elevated expression of the tissue inhibitor of metalloproteinases-1 (TIMP-1) protein and mRNA has been reported in human diseases including cancers and tissue fibrosis.
Regulation of TIMP-1 gene expression is mainly mediated at the level of gene transcription and involves the activation of several well known transcription factors including those belonging to the AP-1, STAT, and Pea3/Ets families. In the current study, we have used DNase-1 footprinting to identify a new regulatory element (5'-TGTGGTTTCCG-3') present in the human TIMP-1 gene promoter. Mutagenesis and transfection studies in culture-activated rat hepatic stellate cells and the human Jurkat T cell line demonstrated that the new element named upstream TIMP-1 element-1 (UTE-1) is essential for transcriptional activity of the human TIMP-1 promoter.
Electrophoretic mobility shift assay studies revealed that UTE-1 can form protein-DNA complexes of distinct mobilities with nuclear extracts from a variety of mammalian cell types and showed that induction of a high mobility UTE-1 complex is associated with culture activation of freshly isolated rat hepatic stellate cells. A combination of UV-cross-linking and Southwestern blotting techniques demonstrated that UTE-1 directly interacts with a 30-kDa nuclear protein that appears to be present in all cell types tested. We conclude that UTE-1 is a novel regulatory element that in combination with its cellular binding proteins may be an important component of the mechanisms controlling TIMP-1 expression in normal and pathological states.
Regulation of TIMP-1 gene expression is mainly mediated at the level of gene transcription and involves the activation of several well known transcription factors including those belonging to the AP-1, STAT, and Pea3/Ets families. In the current study, we have used DNase-1 footprinting to identify a new regulatory element (5'-TGTGGTTTCCG-3') present in the human TIMP-1 gene promoter. Mutagenesis and transfection studies in culture-activated rat hepatic stellate cells and the human Jurkat T cell line demonstrated that the new element named upstream TIMP-1 element-1 (UTE-1) is essential for transcriptional activity of the human TIMP-1 promoter.
Electrophoretic mobility shift assay studies revealed that UTE-1 can form protein-DNA complexes of distinct mobilities with nuclear extracts from a variety of mammalian cell types and showed that induction of a high mobility UTE-1 complex is associated with culture activation of freshly isolated rat hepatic stellate cells. A combination of UV-cross-linking and Southwestern blotting techniques demonstrated that UTE-1 directly interacts with a 30-kDa nuclear protein that appears to be present in all cell types tested. We conclude that UTE-1 is a novel regulatory element that in combination with its cellular binding proteins may be an important component of the mechanisms controlling TIMP-1 expression in normal and pathological states.
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Päivitys 18.4. 2014
Päivitys 18.4. 2014
