B Meyrick and MA Magnuson
Department of Pathology, Vanderbilt University Medical Center, Nashville, TN 36232-2650.

Manganous superoxide dismutase (MnSOD) gene expression is stimulated by endotoxin, tumor necrosis factor, and interleukin-1, agents thought to cause cellular damage through intracellular generation of reactive oxygen species. To study the molecular mechanisms underlying the induction of MnSOD mRNA by these stimuli, we cloned a bovine MnSOD cDNA and used it to isolate the promoter region of the bovine MnSOD gene. A 14 kb genomic DNA fragment (lambda BS1) containing the first and second exons and 5' flanking region of the gene was characterized. The transcription start site was determined by primer extension and S1 nuclease protection assays and found to be 88 bp upstream of the translation initiation codon. The sequence of approximately 1 kb of DNA upstream of the start site was determined and examined for potential regulatory elements. DNA immediately upstream of the transcription start site was GC-rich and contained two AP-2 and eight Sp-1 consensus sequences. It did not contain either a CCAAT or TATA box. A 956 bp fragment of this DNA fragment was transcriptionally active when fused to a luciferase reporter gene and transfected into both bovine pulmonary artery endothelial and hamster insulinoma tumor cells. Transfection analysis of three additional deletion mutants, whose 5' end-points were -317, -182, and -70 bp, respectively, showed a step- like reduction in transfection efficiency, suggesting the presence of regulatory elements throughout this DNA fragment that contribute to transcriptional activity of the MnSOD promoter. Despite the high homology of the bovine MnSOD cDNA to other mammalian MnSODs, the promoter sequences of bovine and rat MnSOD genes showed a virtual lack of similarity.(ABSTRACT TRUNCATED AT 250 WORDS)

This article has been cited by other articles:

				S. K. Dhar, Y. Xu, T. Noel, and D. K. St Clair Chronic exposure to 12-O-tetradecanoylphorbol-13-acetate represses sod2 induction in vivo: the negative role of p50 Carcinogenesis, December 1, 2007; 28(12): 2605 - 2613. [Abstract] [Full Text] [PDF]

				S. K. Dhar, B. C. Lynn, C. Daosukho, and D. K. St. Clair Identification of Nucleophosmin as an NF-{kappa}B Co-activator for the Induction of the Human SOD2 Gene J. Biol. Chem., July 2, 2004; 279(27): 28209 - 28219. [Abstract] [Full Text] [PDF]

				J. B. Wijeweera, A. J. Gandolfi, A. Parrish, and R. C. Lantz Sodium Arsenite Enhances AP-1 and NF{{kappa}} B DNA Binding and Induces Stress Protein Expression in Precision-Cut Rat Lung Slices Toxicol. Sci., June 1, 2001; 61(2): 283 - 294. [Abstract] [Full Text] [PDF]

				H.-P. Kim, J.-H. Roe, P. B. Chock, and M. B. Yim Transcriptional Activation of the Human Manganese Superoxide Dismutase Gene Mediated by Tetradecanoylphorbol Acetate J. Biol. Chem., December 24, 1999; 274(52): 37455 - 37460. [Abstract] [Full Text] [PDF]

				J. Hollander, R. Fiebig, M. Gore, J. Bejma, T. Ookawara, H. Ohno, and L. L. Ji Superoxide dismutase gene expression in skeletal muscle: fiber-specific adaptation to endurance training Am J Physiol Regulatory Integrative Comp Physiol, September 1, 1999; 277(3): R856 - R862. [Abstract] [Full Text] [PDF]

				X. Bu and T. Quertermous Identification of an Endothelial Cell-specific Regulatory Region in the Murine Endothelin-1 Gene J. Biol. Chem., December 19, 1997; 272(51): 32613 - 32622. [Abstract] [Full Text] [PDF]

				S. Leers-Sucheta, K.-i. Morohashi, J. I. Mason, and M. H. Melner Synergistic Activation of the Human Type II 3beta -Hydroxysteroid Dehydrogenase/Delta 5-Delta 4 Isomerase Promoter by the Transcription Factor Steroidogenic Factor-1/Adrenal 4-binding Protein and Phorbol Ester J. Biol. Chem., March 21, 1997; 272(12): 7960 - 7967. [Abstract] [Full Text] [PDF]

				C.-H. Zhu, Y. Huang, L. W. Oberley, and F. E. Domann A Family of AP-2 Proteins Down-regulate Manganese Superoxide Dismutase Expression J. Biol. Chem., April 20, 2001; 276(17): 14407 - 14413. [Abstract] [Full Text] [PDF]