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dc.contributor.authorBhat, Rashid
dc.contributor.authorWeaver, James A.
dc.contributor.authorWagner, Conrad
dc.contributor.authorBodwell, Jack E.
dc.contributor.authorBresnick, Edward
dc.date2022-08-11T08:10:04.000
dc.date.accessioned2022-08-23T16:54:11Z
dc.date.available2022-08-23T16:54:11Z
dc.date.issued1996-12-20
dc.date.submitted2008-08-15
dc.identifier.citation<p>J Biol Chem. 1996 Dec 20;271(51):32551-6.</p>
dc.identifier.issn0021-9258 (Print)
dc.identifier.doi10.1074/jbc.271.51.32551
dc.identifier.pmid8955080
dc.identifier.urihttp://hdl.handle.net/20.500.14038/42443
dc.description.abstractIn the rat, cytochrome P4501A1 gene expression is thought to be regulated by several trans-acting factors including the 4 S polycyclic aromatic hydrocarbon (PAH)-binding protein. Phosphorylation and dephosphorylation have been suggested to influence the function of many cytosolic receptors and transcription factors. The ATP level within H4IIE rat hepatoma cells could be depleted by treatment with sodium azide or 2,4-dinitrophenol; restoration of the original ATP levels occurred with addition of glucose to the cell culture. ATP depletion reduced the phosphate content of the 4 S protein by approximately 25-30%, which lowered the binding of benzo[a]pyrene (B[a]P) to the 4 S protein by >60%. This effect could not be reversed by the addition of ATP to the binding reaction mixtures. Alkaline phosphatase treatment of the purified 4 S protein in a cell-free system also reduced the B[a]P binding to the protein. Cells treated with a protein phosphatase inhibitor, okadaic acid, and a protein kinase inhibitor, staurosporin, affected the B[a]P binding of the 4 S protein positively and negatively, respectively. These data suggested that phosphorylation is involved in the interaction of the 4 S protein with the PAH. The nuclear translocation of the predominantly cytosolic binding protein has been investigated after ligand binding. Western blots with the immunopurified 4 S PAH-binding protein from cytosolic and nuclear lysates showed significant differences in the distribution of the 4 S receptor between cytosolic and nuclear compartments in control and ATP-depleted cells. Ligand binding stimulated the movement of the receptor into the nucleus, which was completely blocked by reducing the intracellular ATP concentration. These findings provide new information on the role of ATP and phosphorylation on the interaction of B[a]P with 4 S PAH-binding protein and its nuclear translocation.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=8955080&dopt=Abstract">Link to Article in PubMed</a></p>
dc.relation.urlhttps://doi.org/10.1074/jbc.271.51.32551
dc.subject2,4-Dinitrophenol
dc.subjectAdenosine Triphosphate
dc.subjectAnimals
dc.subjectAzides
dc.subjectBenzo(a)pyrene
dc.subjectBiological Transport
dc.subjectCarrier Proteins
dc.subjectCell Compartmentation
dc.subjectCell Nucleus
dc.subjectCell Survival
dc.subjectCells, Cultured
dc.subjectCytochrome P-450 CYP1A1
dc.subjectCytosol
dc.subjectGlycine N-Methyltransferase
dc.subjectLiver Neoplasms, Experimental
dc.subject*Methyltransferases
dc.subjectOkadaic Acid
dc.subjectPhosphoprotein Phosphatases
dc.subjectRats
dc.subjectStaurosporine
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleATP depletion affects the phosphorylation state, ligand binding, and nuclear transport of the 4 S polycyclic aromatic hydrocarbon-binding protein in rat hepatoma cells
dc.typeJournal Article
dc.source.journaltitleThe Journal of biological chemistry
dc.source.volume271
dc.source.issue51
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/792
dc.identifier.contextkey579670
html.description.abstract<p>In the rat, cytochrome P4501A1 gene expression is thought to be regulated by several trans-acting factors including the 4 S polycyclic aromatic hydrocarbon (PAH)-binding protein. Phosphorylation and dephosphorylation have been suggested to influence the function of many cytosolic receptors and transcription factors. The ATP level within H4IIE rat hepatoma cells could be depleted by treatment with sodium azide or 2,4-dinitrophenol; restoration of the original ATP levels occurred with addition of glucose to the cell culture. ATP depletion reduced the phosphate content of the 4 S protein by approximately 25-30%, which lowered the binding of benzo[a]pyrene (B[a]P) to the 4 S protein by >60%. This effect could not be reversed by the addition of ATP to the binding reaction mixtures. Alkaline phosphatase treatment of the purified 4 S protein in a cell-free system also reduced the B[a]P binding to the protein. Cells treated with a protein phosphatase inhibitor, okadaic acid, and a protein kinase inhibitor, staurosporin, affected the B[a]P binding of the 4 S protein positively and negatively, respectively. These data suggested that phosphorylation is involved in the interaction of the 4 S protein with the PAH. The nuclear translocation of the predominantly cytosolic binding protein has been investigated after ligand binding. Western blots with the immunopurified 4 S PAH-binding protein from cytosolic and nuclear lysates showed significant differences in the distribution of the 4 S receptor between cytosolic and nuclear compartments in control and ATP-depleted cells. Ligand binding stimulated the movement of the receptor into the nucleus, which was completely blocked by reducing the intracellular ATP concentration. These findings provide new information on the role of ATP and phosphorylation on the interaction of B[a]P with 4 S PAH-binding protein and its nuclear translocation.</p>
dc.identifier.submissionpathoapubs/792
dc.contributor.departmentDepartment of Pharmacology and Molecular Toxicology
dc.source.pages32551-6


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