Regulation of ppp(A2'p)nA-dependent RNase levels during interferon treatment and cell differentiation.

Regulation of ppp(A2'p)nA-dependent RNase levels during interferon treatment and cell differentiation.
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在干扰素治疗和细胞分化过程中调节 ppp(A2p)nA 依赖性 RNase 水平。

DOI:
10.1111/j.1432-1033.1985.tb08695.x
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发表时间:
1985
期刊:
European journal of biochemistry
影响因子:
--
通讯作者:
Friedman,RM
Friedman,RM
中科院分区:
--
文献类型:
--
作者:
Krause,D;Silverman,RH;Jacobsen,H;Leisy,SA;Dieffenbach,CW;Friedman,RM

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细胞内效应寡核苷酸ppp(A2 'p)nA(n= 2-≥4)通过激活ppp(A2' p)nA依赖性RNase来调节RNA的分解。这种RNA酶的细胞水平被证明是在小鼠胚胎癌细胞的分化过程中进行调节。通过分析将ppp(A2′p)nA引入完整细胞后的rRNA分解,在三种分化细胞类型(F9克隆9、PYS和PSA 5E)中证实了干扰素对该RNA酶的诱导。相比之下,在三种未分化胚胎癌细胞系(F9、PC 13克隆5和Nulli 2A)中,无论有无干扰素预处理,几乎没有ppp(A2′p)nA依赖性RNase。这些结果通过无细胞系统中RNA酶的亲和标记得到证实。向细胞裂解缓冲液中加入蛋白酶抑制剂亮抑酶肽是稳定RNA酶以防止其裂解为离散的分解产物所必需的。此外,在视黄酸和N6,O2′-二丁酰腺苷3′,5 ′-单磷酸诱导的PC 13克隆5细胞分化过程中,ppp(A2′p)nA-依赖性RNase逐渐被诱导。因此,这种RNA酶的表达在细胞分化过程中大大增强。此外,研究了双链RNA依赖性蛋白激酶,发现无论细胞分化状态如何,在所有细胞系中均为干扰素诱导型。
The intracellular effector oligonucleotides ppp(A2′p)nA (n= 2−≥4) regulate the breakdown of RNA by activating ppp(A2′p)nA‐dependent RNase. Cellular levels of this RNase were demonstrated to be regulated during differentiation of murine embryonal carcinoma cells. An induction of this RNase by interferon was demonstrated in each of three differentiated cell types (F9 clone 9, PYS, and PSA 5E) by analyzing rRNA breakdown following the introduction of ppp(A2′p)nA into the intact cells. In contrast, in three undifferentiated embryonal carcinoma cell lines (F9, PC13 clone 5, and Nulli 2A) there was little if any ppp(A2′p)nA‐dependent RNase either with or without interferon pretreatment. These results were confirmed by affinity labeling of the RNase in cell‐free systems. Addition of the proteinase inhibitor, leupeptin, to the cell lysis buffer was necessary to stabilize the RNase against cleavage to discrete breakdown products. Moreover, during differentiation of PC13 clone 5 cells by retinoic acid andN6,O2′‐dibutyryl‐adenosine 3′,5′‐monophosphate there was a gradual induction of ppp(A2′p)nA‐dependent RNase. The expression of this RNase is, therefore, greatly enhanced during cell differentiation. In addition, the double‐stranded‐RNA‐dependent protein kinase was investigated and was found to be interferon‐inducible in all of the cell lines regardless of the state of cell differentiation.
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