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正常小鼠胸腺组织中脑池内A颗粒基因的表达:基因产物及品系相关变异性

Intracisternal A-particle gene expression in normal mouse thymus tissue: gene products and strain-related variability.

作者信息

Kuff E L, Fewell J W

出版信息

Mol Cell Biol. 1985 Mar;5(3):474-83. doi: 10.1128/mcb.5.3.474-483.1985.

Abstract

Intracisternal A-particle (IAP)-specific sequences were 5- to 10-fold enriched in polyadenylated RNA from BALB/cJ thymus as compared with RNAs from liver, spleen, and kidney. The major transcripts of 7.2 and 5.4 kilobases were the same size as those found in an IAP-rich neuroblastoma cell line. The absolute levels and proportions of these transcripts varied in thymuses from mice of different inbred strains. With antiserum prepared against p73, the main IAP structural protein, several size classes of IAP-related proteins were immunoprecipitated from extracts of thymus cells incubated with [35S]methionine; these included p73 itself and a group of polypeptides in the size range of 114 to 120 kilodaltons (p114-p120). The inbred strains showed marked characteristic differences in the electrophoretic patterns of their IAP-related proteins. Earlier studies showed that the 7.2-kilobase RNA from neuroblastoma IAPs coded for p73 in a cell-free translation system. Correlations between the RNA and protein patterns in thymuses of the different inbred strains indicated that 5.4-kilobase RNA gives rise to the p114-p120 polypeptides. Metabolically labeled p120 was found to include methionine-containing tryptic peptides of p73 plus additional peptides consistent with its larger size. In vivo labeling kinetics showed that the p114-p120 polypeptides were not major precursors of p73 in intact neuroblastoma cells. This study shows that IAP gene expression in mouse thymus is genetically determined and that a novel class of IAP-related polypeptides can be expressed independently of the major particle structural protein.

摘要

与来自肝脏、脾脏和肾脏的RNA相比,BALB/cJ胸腺的聚腺苷酸化RNA中脑池内A颗粒(IAP)特异性序列富集了5至10倍。7.2和5.4千碱基的主要转录本与富含IAP的神经母细胞瘤细胞系中发现的转录本大小相同。这些转录本的绝对水平和比例在不同近交系小鼠的胸腺中有所不同。用针对主要IAP结构蛋白p73制备的抗血清,从用[35S]甲硫氨酸孵育的胸腺细胞提取物中免疫沉淀出几种大小类别的IAP相关蛋白;这些包括p73本身以及一组大小在114至120千道尔顿范围内的多肽(p114 - p120)。近交系在其IAP相关蛋白的电泳图谱上显示出明显的特征差异。早期研究表明,神经母细胞瘤IAP的7.2千碱基RNA在无细胞翻译系统中编码p73。不同近交系胸腺中RNA和蛋白质模式之间的相关性表明,5.4千碱基RNA产生p114 - p120多肽。发现代谢标记的p120包括p73含甲硫氨酸的胰蛋白酶肽以及与其较大尺寸一致的其他肽。体内标记动力学表明,在完整的神经母细胞瘤细胞中,p114 - p120多肽不是p73的主要前体。这项研究表明,小鼠胸腺中IAP基因表达是由基因决定的,并且一类新的IAP相关多肽可以独立于主要颗粒结构蛋白表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b5d/366739/25498b053695/molcellb00099-0054-a.jpg

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