在氟乳清酸存在的情况下标记的信使核糖核酸与大鼠肝脏内质网膜的直接关联。
Direct association of messenger RNA labeled in the presence of fluoroorotate with membranes of the endoplasmic reticulum in rat liver.
作者信息
Cardelli J, Long B, Pitot H C
出版信息
J Cell Biol. 1976 Jul;70(1):47-58. doi: 10.1083/jcb.70.1.47.
Abstract
Liver rough endoplasmic reticulum (RER) membranes were isolated from rats given [3H]orotic acid for 48 h (ribosomal RNA [rRNA] label) or for 3 h along with 5-fluoroorotate; this latter procedure permits the labeling of cytoplasmic messenger RNAs (mRNAs) in the absence of rRNA labeling. More than 50% of the labeled mRNA remained attached to membranes of the RER after complete removal of ribosomes with a buffer of high ionic strength in the presence of puromycin. Under similar conditions, membranes retained 40% of their polyadenylate as determined by a [3H]-polyuridylate hybridization assay. Treatment of mRNA-labeled endoplasmic reticulum membranes with pancreatic RNase indicates that the polyadenylate and possibly nonpolyadenylate-pyrimidine portions of the messenger are involved in the binding of mRNA to the membranes. The implication of these results in furthering our understanding of the mechanisms of the translational regulation of genetic expression is discussed.
摘要
从给予[3H]乳清酸48小时（核糖体RNA[rRNA]标记）或与5-氟乳清酸一起给予3小时的大鼠中分离肝脏粗面内质网（RER）膜；后一种方法允许在不存在rRNA标记的情况下标记细胞质信使RNA（mRNA）。在用高离子强度缓冲液在嘌呤霉素存在下完全去除核糖体后，超过50%的标记mRNA仍附着在RER膜上。在类似条件下，通过[3H]-聚尿苷酸杂交测定法确定，膜保留了其40%的聚腺苷酸。用胰腺核糖核酸酶处理mRNA标记的内质网膜表明，信使的聚腺苷酸部分以及可能的非聚腺苷酸-嘧啶部分参与了mRNA与膜的结合。讨论了这些结果对深化我们对基因表达翻译调控机制理解的意义。
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