Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.11.2 (PDK1)
 2,238 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In the present study the hepatitis B virus X antigen binding protein 1 (XBP1) was cloned by inducing its expression, and its subcellular localization and function were examined. Total RNA was extracted from HepG2 cells and XBP1 was amplified using reverse transcription polymerase chain reaction (RT-PCR), followed by restriction enzyme digestion of the pGBKT7 yeast plasmid and identification by enzyme digestion. The plasmid was transformed into AH109 yeast via the lithium acetate method and protein extracts were prepared. XBP1 protein expression in the eukaryotic cells was determined using polyacrylamide gel electrophoresis and western blot analysis. The gene encoding the XBP1-binding protein was screened in liver cells using yeast two-hybrid technology. We transfected a human hepatocellular carcinoma cell line and observed the intracellular localization of the gene expression protein using a fluorescence microscope, followed by prokaryotic expression and XBP1 gene identification. A 921-bp XBP1 gene fragment was obtained via RT-PCR amplification and 20 proteins with known functions that interact with XBP1 were screened, including metallothionein, smooth muscle cell-related protein, asialoglycoprotein receptor, pyruvate dehydrogenase kinase 1 and a sequence with unknown functions. A green fluorescent protein expression plasmid pEGFP-C1-XBP1 of XBP1 was constructed successfully and its expression protein was localized in the cytoplasm. A 56-kDa recombinant protein was successfully obtained via prokaryotic expression and was demonstrated to have good specificity using western blot analysis. The XBP1 gene, which expresses the XBP1 protein, is located in the cytoplasm and plays a role in the intracellular structure, cell growth, intracellular metabolism and signal transduction pathway, as well as DNA duplication, transcription, recombination and repair.
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PMID:Cloning and functions of the HBxAg-binding protein XBP1. 2324 34

It has been proposed that aging results from the lifelong accumulation of intracellular damage via reactions with reactive oxygen species (ROS). Metallothioneins are conserved cysteine-rich proteins that function as efficient ROS scavengers and may affect longevity. To better understand mechanisms controlling metallothionein expression, the regulatory factors and pathways that controlled cadmium-inducible transcription of the C. elegans metallothionein gene, mtl-1, were identified. The transcription factor ATF-7 was identified in both ethylmethanesulfonate mutagenesis and candidate gene screens. PMK-1 and members of the insulin signaling pathway, PDK-1 and AKT-1/2, were also identified as mtl-1 regulators. Genetic and previous results support a model for the regulation of cadmium-inducible mtl-1 transcription based on the derepression of the constitutively active transcription factor ELT-2. In addition, knockdown of the mammalian homologs of PDK1 and ATF7 in HEK293 cells resulted in changes in metallothionein expression, suggesting that this pathway was evolutionarily conserved. The insulin signaling pathway is known to influence the aging process; however, various factors responsible for affecting the aging phenotype are unknown. Identification of portions of the insulin signaling pathway as regulators of metallothionein expression supports the hypothesis that longevity is affected by the expression of this efficient ROS scavenger.
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PMID:Identification of ATF-7 and the insulin signaling pathway in the regulation of metallothionein in C. elegans suggests roles in aging and reactive oxygen species. 2863 56