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Sp1/Sp3 and the myeloid zinc finger gene MZF1 regulate the human N-cadherin promoter in osteoblasts.
Exp Cell Res. 2005 Jan 01; 302(1):129-42.EC

Abstract

To determine the molecular mechanisms by which N-cadherin transcription is regulated, we cloned and sequenced a 3681-bp of the 5'-flanking region of the human N-cadherin gene. Deletion analysis of the proximal region identified a minimal 318-bp region with strong promoter activity in human osteoblasts. The cryptic promoter is characterized by high GC content and a GA-rich binding core that may bind zing finger transcription factors. Electrophoretic mobility shift assays (EMSA), competition and supershift EMSA revealed that an Sp1/Sp3 binding site acts as a basal regulatory element of the promoter in osteoblasts. Incubation of osteoblast nuclear extracts with -163/-131 wild-type probe containing the GA-rich binding core revealed another specific complex, which was not formed with a -163/-131 probe mutated in the GA repeat. EMSA identified the nuclear factor involved as myeloid zinc finger-1 (MZF1). Mutation analysis showed that Sp1/Sp3 and MZF1 binding sites contribute to basal promoter activity. Cotransfection analyses showed that Sp1 and MZF1 overexpression increases whereas Sp3 antagonizes Sp1-induced N-cadherin promoter activity in osteoblasts. RT-PCR analysis showed that human osteoblastic cells express MZF1 and that Sp1/MZF1 overexpression increased N-cadherin expression. These results indicate that Sp1/Sp3 and MZF1 are important transcription factors regulating N-cadherin promoter activity and expression in osteoblasts.

Authors+Show Affiliations

Laboratory of Osteoblast Biology and Pathology, INSERM U606, Lariboisière Hospital, 75475 Cedex 10 Paris, France.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15541732

Citation

Le Mée, S, et al. "Sp1/Sp3 and the Myeloid Zinc Finger Gene MZF1 Regulate the Human N-cadherin Promoter in Osteoblasts." Experimental Cell Research, vol. 302, no. 1, 2005, pp. 129-42.
Le Mée S, Fromigué O, Marie PJ. Sp1/Sp3 and the myeloid zinc finger gene MZF1 regulate the human N-cadherin promoter in osteoblasts. Exp Cell Res. 2005;302(1):129-42.
Le Mée, S., Fromigué, O., & Marie, P. J. (2005). Sp1/Sp3 and the myeloid zinc finger gene MZF1 regulate the human N-cadherin promoter in osteoblasts. Experimental Cell Research, 302(1), 129-42.
Le Mée S, Fromigué O, Marie PJ. Sp1/Sp3 and the Myeloid Zinc Finger Gene MZF1 Regulate the Human N-cadherin Promoter in Osteoblasts. Exp Cell Res. 2005 Jan 1;302(1):129-42. PubMed PMID: 15541732.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Sp1/Sp3 and the myeloid zinc finger gene MZF1 regulate the human N-cadherin promoter in osteoblasts. AU - Le Mée,S, AU - Fromigué,O, AU - Marie,P J, PY - 2004/04/14/received PY - 2004/08/19/revised PY - 2004/11/16/pubmed PY - 2005/1/14/medline PY - 2004/11/16/entrez SP - 129 EP - 42 JF - Experimental cell research JO - Exp Cell Res VL - 302 IS - 1 N2 - To determine the molecular mechanisms by which N-cadherin transcription is regulated, we cloned and sequenced a 3681-bp of the 5'-flanking region of the human N-cadherin gene. Deletion analysis of the proximal region identified a minimal 318-bp region with strong promoter activity in human osteoblasts. The cryptic promoter is characterized by high GC content and a GA-rich binding core that may bind zing finger transcription factors. Electrophoretic mobility shift assays (EMSA), competition and supershift EMSA revealed that an Sp1/Sp3 binding site acts as a basal regulatory element of the promoter in osteoblasts. Incubation of osteoblast nuclear extracts with -163/-131 wild-type probe containing the GA-rich binding core revealed another specific complex, which was not formed with a -163/-131 probe mutated in the GA repeat. EMSA identified the nuclear factor involved as myeloid zinc finger-1 (MZF1). Mutation analysis showed that Sp1/Sp3 and MZF1 binding sites contribute to basal promoter activity. Cotransfection analyses showed that Sp1 and MZF1 overexpression increases whereas Sp3 antagonizes Sp1-induced N-cadherin promoter activity in osteoblasts. RT-PCR analysis showed that human osteoblastic cells express MZF1 and that Sp1/MZF1 overexpression increased N-cadherin expression. These results indicate that Sp1/Sp3 and MZF1 are important transcription factors regulating N-cadherin promoter activity and expression in osteoblasts. SN - 0014-4827 UR - https://www.unboundmedicine.com/medline/citation/15541732/Sp1/Sp3_and_the_myeloid_zinc_finger_gene_MZF1_regulate_the_human_N_cadherin_promoter_in_osteoblasts_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0014-4827(04)00507-5 DB - PRIME DP - Unbound Medicine ER -