Chan Ching Wan, Lee Youn-Bok, Uney James, Flynn Andrea, Tobias Jonathan H, Norman Michael
Henry Wellcome Laboratories for Integrative Neurosciences and Endocrinology, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK.
Biochem J. 2007 Nov 1;407(3):355-62. doi: 10.1042/BJ20070170.
The SLTM [SAF (scaffold attachment factor)-like transcription modulator] protein contains a SAF-box DNA-binding motif and an RNA-binding domain, and shares an overall identity of 34% with SAFB1 {scaffold attachment factor-B1; also known as SAF-B (scaffold attachment factor B), HET [heat-shock protein 27 ERE (oestrogen response element) and TATA-box-binding protein] or HAP (heterogeneous nuclear ribonucleoprotein A1-interacting protein)}. Here, we show that SLTM is localized to the cell nucleus, but excluded from nucleoli, and to a large extent it co-localizes with SAFB1. In the nucleus, SLTM has a punctate distribution and it does not co-localize with SR (serine/arginine) proteins. Overexpression of SAFB1 has been shown to exert a number of inhibitory effects, including suppression of oestrogen signalling. Although SLTM also suppressed the ability of oestrogen to activate a reporter gene in MCF-7 breast-cancer cells, inhibition of a constitutively active beta-galactosidase gene suggested that this was primarily the consequence of a generalized inhibitory effect on transcription. Measurement of RNA synthesis, which showed a particularly marked inhibition of [(3)H]uridine incorporation into mRNA, supported this conclusion. In addition, analysis of cell-cycle parameters, chromatin condensation and cytochrome c release showed that SLTM induced apoptosis in a range of cultured cell lines. Thus the inhibitory effects of SLTM on gene expression appear to result from generalized down-regulation of mRNA synthesis and initiation of apoptosis consequent upon overexpressing the protein. While indicating a crucial role for SLTM in cellular function, these results also emphasize the need for caution when interpreting phenotypic changes associated with manipulation of protein expression levels.
SLTM[类SAF(支架附着因子)转录调节因子]蛋白包含一个SAF框DNA结合基序和一个RNA结合结构域,与SAFB1{支架附着因子-B1;也称为SAF-B(支架附着因子B)、HET[热休克蛋白27雌激素反应元件和TATA框结合蛋白]或HAP(异质性核糖核蛋白A1相互作用蛋白)}的总体一致性为34%。在此,我们表明SLTM定位于细胞核,但排除在核仁之外,并且在很大程度上与SAFB1共定位。在细胞核中,SLTM呈点状分布,并且不与SR(丝氨酸/精氨酸)蛋白共定位。SAFB1的过表达已显示出多种抑制作用,包括抑制雌激素信号传导。虽然SLTM也抑制雌激素激活MCF-7乳腺癌细胞中报告基因的能力,但对组成型活性β-半乳糖苷酶基因的抑制表明,这主要是对转录的普遍抑制作用的结果。RNA合成的测量显示[(3)H]尿苷掺入mRNA受到特别明显的抑制,支持了这一结论。此外,细胞周期参数、染色质凝聚和细胞色素c释放的分析表明,SLTM在一系列培养细胞系中诱导细胞凋亡。因此,SLTM对基因表达的抑制作用似乎是由于mRNA合成的普遍下调以及过表达该蛋白后引发的细胞凋亡。这些结果虽然表明SLTM在细胞功能中起关键作用,但也强调在解释与蛋白质表达水平操作相关的表型变化时需要谨慎。