Nuclear protein that binds sterol regulatory element of low density lipoprotein receptor promoter. II. Purification and characterization

作者：Xiaodong Wang, Michael R. Briggs, Xianxin Hua, Chieko Yokoyama, J L Goldstein, Michael S. Brown · 发表于：Journal of Biological Chemistry · 年份：1993 · DOI：10.1016/s0021-9258(19)85266-3 · 被引用次数：301 · 研究领域：Cholesterol and Lipid Metabolism、RNA Research and Splicing、Cell Adhesion Molecules Research

This paper describes the purification and characterization of a sterol regulatory element binding protein (SREBP) that recognizes the SRE-1 sequence in the 5' flanking region of the gene for the low density lipoprotein (LDL) receptor. The protein was purified more than 38,000-fold from nuclear extracts of human HeLa cells by ion exchange, gel filtration, and DNA-affinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified preparation revealed a cluster of bands at 59-68 kDa, each of which bound to the SRE-1 element as revealed by cross-linking experiments. Binding of SREBP correlated perfectly with transcriptional activity in a series of 16 sterol regulatory elements with point mutations. In the LDL receptor promoter, the 10-base pair SRE-1 is embedded in a 16-base pair sequence designated Repeat 2, which is adjacent to Repeat 3, a binding site for nuclear factor Sp1. Oligonucleotides containing Repeat 2 + 3 bound SREBP and Sp1 as revealed by mobility shift assays. SREBP produced a DNase I footprint over the SRE-1 sequence, which was immediately adjacent to the footprint produced by Sp1. The current data are consistent with the concept that SREBP acts in concert with Sp1 to achieve high level, sterol-suppressible transcription of the gene for the LDL receptor.