Sasahara M, Amano S, Sato H, Yang J G, Hayase Y, Kaneko M, Sato I, Suzaki M, Hazama F
Department of Pathology, Shiga University of Medical Sciences, Otsu City, Japan.
Oncogene. 1998 Mar 26;16(12):1571-8. doi: 10.1038/sj.onc.1201679.
The 5' untranslated sequence (5' UTS) of platelet-derived growth factor B (PDGF-B/c-sis) mRNA is highly preserved through evolution, and inhibits translation of downstream coding sequences. In this study, using Northern analysis we identified two PDGF-B/c-sis mRNAs (3.5 kb and 2.6 kb) expressed in normal developing rat brain. In contrast to the constitutive expression of 3.5 kb mRNA, the expression of 2.6 kb mRNA increased markedly in accordance with those stages of brain development at which we had previously demonstrated an increased immunoreactivity for PDGF-B/c-SIS in neurons (Sasahara et al., 1992). By PCR cloning and the RNase protection assay, we determined the complete sequence of rat PDGF-B/c-sis, and found that the 2.6 kb transcript was a form of the 3.5 kb message truncated at the 5' end, and that the predominant 2.6 kb mRNA commenced 15 nt upstream of the signal peptide. Accordingly, it is suggested that the truncation of 5' UTS contributes to the expression of PDGF-B/c-SIS protein in the CNS. Lack of translational inhibitory 5' UTS of PDGF-B/c-sis transcript and resultant efficient protein translation have been reported in only a few transformed cells and cultured umbilical vein endothelial cells. We have extended this knowledge to the developing rat brain, and suggest that a similar mechanism could operate widely in non-transformed tissue in vivo.
血小板衍生生长因子B(PDGF - B/c - sis)mRNA的5'非翻译序列(5' UTS)在进化过程中高度保守,并抑制下游编码序列的翻译。在本研究中,我们通过Northern分析鉴定出在正常发育的大鼠脑中表达的两种PDGF - B/c - sis mRNA(3.5 kb和2.6 kb)。与3.5 kb mRNA的组成性表达不同,2.6 kb mRNA的表达随着脑发育阶段的进展而显著增加,而我们之前已证明在这些阶段神经元中PDGF - B/c - SIS的免疫反应性增强(Sasahara等人,1992年)。通过PCR克隆和核糖核酸酶保护试验,我们确定了大鼠PDGF - B/c - sis的完整序列,发现2.6 kb转录本是5'端截短的3.5 kb信息的一种形式,并且主要的2.6 kb mRNA在信号肽上游15个核苷酸处起始。因此,提示5' UTS的截短有助于PDGF - B/c - SIS蛋白在中枢神经系统中的表达。仅在少数转化细胞和培养的脐静脉内皮细胞中报道了PDGF - B/c - sis转录本缺乏翻译抑制性5' UTS以及由此产生的高效蛋白质翻译。我们将这一知识扩展到发育中的大鼠脑,并提示类似的机制可能在体内未转化组织中广泛起作用。