Characterization of the neuropilin-1 promoter; Gene expression is mediated by the transcription factor Sp1

Abstract

Neuropilin-1 (NRP1) is a receptor for the vascular endothelial growth factor (VEGF) family of angiogenesis factors and for the semaphorin family of secreted neuronal guidance polypeptides. Very little is known, however, about how NRP1 gene expression is regulated. In this study, it was demonstrated that the tumor promoter, TPA (12-O-tetradecanoylphorbol-13-acetate) significantly up-regulated NRP1 mRNA levels by increasing its gene transcription rate in a manner dependent on de novo protein synthesis. To determine which elements regulate functional NRP1 expression, the promoter regions of human and mouse NRP1 genes were cloned and characterized. Promoter-reporter gene transfection experiments using deletion and point mutations demonstrated that two Sp1 elements are major contributors to both the constitutive and TPA-induced activity of the NRP1 promoter. Gel shift analysis showed a specific binding of the Sp1 transcription factor to those elements. Further mutational analysis revealed that an AP-1, and a CCAAT box also contributed to NRP1 constitutive and TPA-induced promoter activity. It was concluded that NRP1 expression is regulated by the cooperation of several regulatory elements including AP-1, Sp1, and a CCAAT box. © 2003 Wiley-Liss, Inc.