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Involvement of specific proteins (Sp1/Sp3) and nuclear factor Y in basal transcription of the distal promoter of the rat pyruvate carboxylase gene in beta-cells.
Biochem Biophys Res Commun. 2005 Apr 01; 329(1):188-96.BB

Abstract

Pyruvate carboxylase plays diverse roles in different biosynthetic pathways, including glucose-induced insulin secretion in pancreatic beta-cells. We have localized the control region of the P2 promoter by generating a series of 5'-nested deletion constructs, and both 25- and 9-bp internal deletion constructs, as well as by performing site-directed mutagenesis. Transient transfections of these constructs into INS-1 cells identified a CCAAT box and a GC box that are located at -65/-61 and -48/-41, respectively, as the important determinants. Disruption of the GC box resulted in a 4-fold reduction of the reporter activity, while disruption of the proximal CCAAT box (-65/-61) but not the distal CCAAT box (-95/-91) increased the reporter activity by 3-fold. Simultaneous disruptions of both the GC box and the CCAAT box reduced the reporter activity to a level that was close to that of the single GC box mutation. Electrophoretic mobility shift assays (EMSAs) and supershift EMSAs using nuclear extract from INS-1 cells demonstrated that Sp1 and Sp3 bind a GC box while the nuclear factor Y was shown to bind the proximal but not the distal CCAAT box.

Authors+Show Affiliations

Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15721292

Citation

Sunyakumthorn, Piyanate, et al. "Involvement of Specific Proteins (Sp1/Sp3) and Nuclear Factor Y in Basal Transcription of the Distal Promoter of the Rat Pyruvate Carboxylase Gene in Beta-cells." Biochemical and Biophysical Research Communications, vol. 329, no. 1, 2005, pp. 188-96.
Sunyakumthorn P, Boonsaen T, Boonsaeng V, et al. Involvement of specific proteins (Sp1/Sp3) and nuclear factor Y in basal transcription of the distal promoter of the rat pyruvate carboxylase gene in beta-cells. Biochem Biophys Res Commun. 2005;329(1):188-96.
Sunyakumthorn, P., Boonsaen, T., Boonsaeng, V., Wallace, J. C., & Jitrapakdee, S. (2005). Involvement of specific proteins (Sp1/Sp3) and nuclear factor Y in basal transcription of the distal promoter of the rat pyruvate carboxylase gene in beta-cells. Biochemical and Biophysical Research Communications, 329(1), 188-96.
Sunyakumthorn P, et al. Involvement of Specific Proteins (Sp1/Sp3) and Nuclear Factor Y in Basal Transcription of the Distal Promoter of the Rat Pyruvate Carboxylase Gene in Beta-cells. Biochem Biophys Res Commun. 2005 Apr 1;329(1):188-96. PubMed PMID: 15721292.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Involvement of specific proteins (Sp1/Sp3) and nuclear factor Y in basal transcription of the distal promoter of the rat pyruvate carboxylase gene in beta-cells. AU - Sunyakumthorn,Piyanate, AU - Boonsaen,Thirajit, AU - Boonsaeng,Vichai, AU - Wallace,John C, AU - Jitrapakdee,Sarawut, PY - 2004/12/23/received PY - 2005/2/22/pubmed PY - 2005/4/14/medline PY - 2005/2/22/entrez SP - 188 EP - 96 JF - Biochemical and biophysical research communications JO - Biochem Biophys Res Commun VL - 329 IS - 1 N2 - Pyruvate carboxylase plays diverse roles in different biosynthetic pathways, including glucose-induced insulin secretion in pancreatic beta-cells. We have localized the control region of the P2 promoter by generating a series of 5'-nested deletion constructs, and both 25- and 9-bp internal deletion constructs, as well as by performing site-directed mutagenesis. Transient transfections of these constructs into INS-1 cells identified a CCAAT box and a GC box that are located at -65/-61 and -48/-41, respectively, as the important determinants. Disruption of the GC box resulted in a 4-fold reduction of the reporter activity, while disruption of the proximal CCAAT box (-65/-61) but not the distal CCAAT box (-95/-91) increased the reporter activity by 3-fold. Simultaneous disruptions of both the GC box and the CCAAT box reduced the reporter activity to a level that was close to that of the single GC box mutation. Electrophoretic mobility shift assays (EMSAs) and supershift EMSAs using nuclear extract from INS-1 cells demonstrated that Sp1 and Sp3 bind a GC box while the nuclear factor Y was shown to bind the proximal but not the distal CCAAT box. SN - 0006-291X UR - https://www.unboundmedicine.com/medline/citation/15721292/Involvement_of_specific_proteins__Sp1/Sp3__and_nuclear_factor_Y_in_basal_transcription_of_the_distal_promoter_of_the_rat_pyruvate_carboxylase_gene_in_beta_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-291X(05)00171-3 DB - PRIME DP - Unbound Medicine ER -