P53 IS COVALENTLY LINKED TO 5.8S RIBOSOMAL-RNA
P53 IS COVALENTLY LINKED TO 5.8S RIBOSOMAL-RNA
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DOI：
10.1128/mcb.12.11.5145
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发表时间：
1992-11-01
影响因子：
5.3
通讯作者：
CARROLL, RB
中科院分区：
文献类型：
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作者：
FONTOURA, BMA;SOROKINA, EA;CARROLL, RB
我们在此报道了特异性免疫共沉淀与肿瘤抑制基因产物P53相连的RNA的分离和鉴定。经蛋白酶K处理后，P53的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)产生一个单一的、离散的157个核苷酸的RNA，该RNA被克隆、测序，并被鉴定为5.8S rRNA。5.8S rRNA只有在P53蛋白酶切后才能获得。在SDS-PAGE中，游离的5.8S rRNA未与P53共存。这种RNA只从含有p53的细胞中免疫沉淀出来。将P53蛋白裂解得到的无蛋白RNA与含有5.8S rRNA反义序列的单链DNA载体杂交。P53-5.8S rRNA连接的共价性被证明如下：(I)P53和连接的5.8S rRNA在SDS-PAGE中迁移；(Ii)只有在用蛋白酶K处理P53-RNA复合体后，5.8S rRNA才与P53连接的5.8S rRNA迁移不同；以及(Iii)分离的RNA与磷酸丝氨酸连接，推测在5‘端。与单个特异性RNA的共价连接表明，p53可能参与了5.8S rRNA的表达或功能的调节。
We report here the isolation and identification of the RNA specifically immunoprecipitated and covalently linked to the tumor suppressor gene product p53. After treatment with proteinase K, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) band of p53 yields a single, discrete 157-nucleotide RNA, which was cloned, sequenced, and identified as 5.8S rRNA. 5.8S rRNA was obtained only after proteolysis of the p53 SDS-PAGE band. Free 5.8S rRNA did not comigrate with p53 in SDS-PAGE. This RNA was only immunoprecipitated from cells containing p53. Protein-free RNA obtained by proteolysis of the p53 band hybridized to the single-stranded DNA vector containing the antisense sequence of 5.8S rRNA. The covalence of the p53-5.8S rRNA linkage was demonstrated by the following findings: (i) p53 and the linked 5.8S rRNA comigrated in SDS-PAGE; (ii) only after treatment of the p53-RNA complex with proteinase K did the 5.8S rRNA migrate differently from p53-linked 5.8S rRNA; and (iii) this isolated RNA was found linked to phosphoserine, presumably at the 5' end. Covalent linkage to the single, specific RNA suggests that p53 may be involved in regulating the expression or function of 5.8S rRNA.