Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.

Pyruvate carboxylase (PC) is an anaplerotic enzyme that regulates glucose-induced insulin secretion in pancreatic islets. Dysregulation of its expression is associated with type 2 diabetes. Herein we describe the molecular mechanism underlying the glucose-mediated transcriptional regulation of the P...

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Autores principales:	Apilak Wutthisathapornchai, Tuangtong Vongpipatana, Sureeporn Muangsawat, Thirajit Boonsaen, Michael J MacDonald, Sarawut Jitrapakdee
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/507c53d032a44b94be7327a9703dc941
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spelling	oai:doaj.org-article:507c53d032a44b94be7327a9703dc9412021-11-25T06:07:24ZMultiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.1932-620310.1371/journal.pone.0102730https://doaj.org/article/507c53d032a44b94be7327a9703dc9412014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25054881/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Pyruvate carboxylase (PC) is an anaplerotic enzyme that regulates glucose-induced insulin secretion in pancreatic islets. Dysregulation of its expression is associated with type 2 diabetes. Herein we describe the molecular mechanism underlying the glucose-mediated transcriptional regulation of the PC gene. Incubation of the rat insulin cell line INS-1 832/13 with glucose resulted in a 2-fold increase in PC mRNA expression. Transient transfections of the rat PC promoter-luciferase reporter construct in the above cell line combined with mutational analysis indicated that the rat PC gene promoter contains the glucose-responsive element (GRE), comprising three canonical E-boxes (E1, E3 and E4) and one E-box-like element (E2) clustering between nucleotides -546 and -399, upstream of the transcription start site. Mutation of any of these E-boxes resulted in a marked reduction of glucose-mediated transcriptional induction of the reporter gene. Electrophoretic mobility shift assays revealed that the upstream stimulatory factors 1 and 2 (USF1 and USF2) bind to E1, the Specificity Protein-1 (Sp1) binds to E2, USF2 and the carbohydrate responsive element binding protein (ChREBP) binds to E4, while unknown factors binds to E3. High glucose promotes the recruitment of Sp1 to E2 and, USF2 and ChREBP to E4. Silencing the expression of Sp1, USF2 and ChREBP by their respective siRNAs in INS-1 832/13 cells blunted glucose-induced expression of endogenous PC. We conclude that the glucose-mediated transcriptional activation of the rat PC gene is regulated by at least these three transcription factors.Apilak WutthisathapornchaiTuangtong VongpipatanaSureeporn MuangsawatThirajit BoonsaenMichael J MacDonaldSarawut JitrapakdeePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 7, p e102730 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Apilak Wutthisathapornchai Tuangtong Vongpipatana Sureeporn Muangsawat Thirajit Boonsaen Michael J MacDonald Sarawut Jitrapakdee Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.
description	Pyruvate carboxylase (PC) is an anaplerotic enzyme that regulates glucose-induced insulin secretion in pancreatic islets. Dysregulation of its expression is associated with type 2 diabetes. Herein we describe the molecular mechanism underlying the glucose-mediated transcriptional regulation of the PC gene. Incubation of the rat insulin cell line INS-1 832/13 with glucose resulted in a 2-fold increase in PC mRNA expression. Transient transfections of the rat PC promoter-luciferase reporter construct in the above cell line combined with mutational analysis indicated that the rat PC gene promoter contains the glucose-responsive element (GRE), comprising three canonical E-boxes (E1, E3 and E4) and one E-box-like element (E2) clustering between nucleotides -546 and -399, upstream of the transcription start site. Mutation of any of these E-boxes resulted in a marked reduction of glucose-mediated transcriptional induction of the reporter gene. Electrophoretic mobility shift assays revealed that the upstream stimulatory factors 1 and 2 (USF1 and USF2) bind to E1, the Specificity Protein-1 (Sp1) binds to E2, USF2 and the carbohydrate responsive element binding protein (ChREBP) binds to E4, while unknown factors binds to E3. High glucose promotes the recruitment of Sp1 to E2 and, USF2 and ChREBP to E4. Silencing the expression of Sp1, USF2 and ChREBP by their respective siRNAs in INS-1 832/13 cells blunted glucose-induced expression of endogenous PC. We conclude that the glucose-mediated transcriptional activation of the rat PC gene is regulated by at least these three transcription factors.
format	article
author	Apilak Wutthisathapornchai Tuangtong Vongpipatana Sureeporn Muangsawat Thirajit Boonsaen Michael J MacDonald Sarawut Jitrapakdee
author_facet	Apilak Wutthisathapornchai Tuangtong Vongpipatana Sureeporn Muangsawat Thirajit Boonsaen Michael J MacDonald Sarawut Jitrapakdee
author_sort	Apilak Wutthisathapornchai
title	Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.
title_short	Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.
title_full	Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.
title_fullStr	Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.
title_full_unstemmed	Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.
title_sort	multiple e-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/507c53d032a44b94be7327a9703dc941
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Multiple E-boxes in the distal promoter of the rat pyruvate carboxylase gene function as a glucose-responsive element.

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