Ory with the NO. 401 Hospital of the PLA for delivering us the precious assist. We also want to thank Yong WANG, Qian SUN, Yongjian SHI, Hui Zhao, Daoyan WANG and Zhaoyan CHEN for their useful assist in our experiment.Author ContributionsConceived and designed the experiments: PW FA ML. Performed the experiments: PW JL BZ PL LL. Analyzed the information: PL XZ LZ. Contributed reagents/materials/analysis tools: ML. Wrote the paper: PL FA ML.
THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 288, NO. 33, pp. 23943?3952, August 16, 2013 ?2013 by The American Society for Biochemistry and Molecular Biology, Inc. Published inside the U.S.A.Transcription Components Sp1 and Hif2 Mediate Induction on the Copper-transporting ATPase (Atp7a) Gene in Intestinal Epithelial Cells in the course of Hypoxia*SReceived for publication, May possibly 29, 2013, and in revised kind, June 27, 2013 Published, JBC Papers in Press, June 28, 2013, DOI 10.1074/jbc.M113.Liwei Xie and James F. Collins1 From the Food Science and Human Nutrition Division, University of Florida, Gainesville, FloridaBackground: A hypoxia-inducible transcription factor (Hif2 ) mediates induction of intestinal iron and copper transporters through iron deficiency. Outcomes: Specificity factor 1 (Sp1) is necessary for transcriptional induction of an intestinal copper transporter (Atp7a) by Hif2 . Conclusion: Sp1 and Hif2 might synergistically mediate the genetic response to iron deficiency. Significance: Understanding molecular mechanisms governing iron absorption may perhaps enable modulation of this course of action in the course of illness states. Genes with G/C-rich promoters had been up-regulated in the duodenal epithelium of iron-deficient rats such as these encoding iron (e.g. Dmt1 and Dcytb) and copper (e.g. Atp7a and Mt1) metabolism-related proteins. It was shown previously that an intestinal copper transporter (Atp7a) was co-regulated with iron transport-related genes by a hypoxia-inducible transcription factor, Hif2 .Formula of 3-(Difluoromethyl)aniline Within the present study, we sought to test the part of Sp1 in transcriptional regulation of Atp7a expression in the course of iron deprivation/hypoxia. Initial studies in IEC-6 cells showed that mithramycin, an Sp1 inhibitor, lowered expression of Atp7a and iron transport-related genes (Dmt1, Dcytb, and Fpn1) and blocked their induction by CoCl2, a hypoxia mimetic. Consistent with this, overexpression of Sp1 enhanced endogenous Atp7a mRNA and protein expression and stimulated Atp7a, Dmt1, and Dcytb promoter activity. Site-directed mutagenesis and functional analysis of a basal Atp7a promoter construct revealed four functional Sp1 binding web sites that were vital for Hif2 -mediated induction of promoter activity. Furthermore, chromatin immunoprecipitation (ChIP) assays confirmed that Sp1 particularly interacts together with the Atp7a promoter in IEC-6 cells and in rat duodenal enterocytes.Methyl 5-(bromomethyl)picolinate site This investigation has thus revealed a novel aspect of Atp7a gene regulation in which Sp1 may perhaps be vital for the HIF-mediated induction of gene transcription through iron deficiency/hypoxia.PMID:33632031 Understanding regulation of Atp7a expression may well aid additional clarify the physiological part of copper within the upkeep of iron homeostasis. In addition, this Sp1/Hif2 regulatory mechanism might have broader implications for understanding the genetic response of the intestinal epithelium to retain whole-body iron homeostasis throughout states of deficiency.Iron is essential for life since it plays critical roles in biological systems such as those associated to mitochondrial electrontransport and ene.
