Tan Keith, Shaw Anthony L, Madsen Bente, Jensen Kirsten, Taylor-Papadimitriou Joyce, Freemont Paul S
Centre for Structural Biology, Department of Biological Sciences, Imperial College London, Armstrong Road, London SW7 2AZ.
J Biol Chem. 2003 Jun 6;278(23):20507-13. doi: 10.1074/jbc.M301994200. Epub 2003 Mar 25.
PLU-1 is a large (1544 amino acids) nuclear protein that is highly expressed in breast cancers and is proposed to function as a regulator of gene expression. A yeast two-hybrid screen using PLU-1 as bait has identified two unrelated PLU-1 interacting proteins, namely brain factor-1 (BF-1) and paired box 9 (PAX9), both of which are developmental transcription factors. BF-1 and PAX9 interact with PLU-1 via a novel conserved sequence motif (Ala-X-Ala-Ala-X-Val-Pro-X4-Val-Pro-X8-Pro, termed the VP motif), because deletion or site-directed mutagenesis of this motif in either protein abolishes PLU-1 interaction in vivo. In a reporter assay system, PLU-1 has potent transcriptional repression activity. BF-1 and PAX9 also represses transcription in the same assay, but co-expression of PLU-1 with BF-1 or PAX9 significantly enhances this repression. Mutation of the PLU-1 binding motifs in BF-1 and PAX9 abolishes the observed PLU-1 co-repression activity. These data support a role for PLU-1 acting as a transcriptional co-repressor of two unrelated developmental transcription factors. Because both BF-1 and PAX proteins interact with members of the groucho co-repressor family, it is plausible that PLU-1 has a role in groucho-mediated transcriptional repression.
PLU-1是一种大型(1544个氨基酸)核蛋白,在乳腺癌中高度表达,被认为具有基因表达调节因子的功能。以PLU-1为诱饵进行的酵母双杂交筛选鉴定出两种与PLU-1相互作用的不相关蛋白,即脑因子-1(BF-1)和配对盒9(PAX9),它们都是发育转录因子。BF-1和PAX9通过一个新的保守序列基序(Ala-X-Ala-Ala-X-Val-Pro-X4-Val-Pro-X8-Pro,称为VP基序)与PLU-1相互作用,因为这两种蛋白中该基序的缺失或定点诱变都会消除体内PLU-1的相互作用。在报告基因检测系统中,PLU-1具有强大的转录抑制活性。BF-1和PAX9在同一检测中也能抑制转录,但PLU-与BF-1或PAX9共表达会显著增强这种抑制作用。BF-1和PAX9中PLU-1结合基序的突变消除了观察到的PLU-1共抑制活性。这些数据支持PLU-1作为两种不相关发育转录因子的转录共抑制因子的作用。由于BF-1和PAX蛋白都与gro同家族的成员相互作用,因此PLU-1在gro介导的转录抑制中发挥作用是合理的。