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棘阿米巴中核糖体成分合成的协调调控:在包囊形成过程中,5S RNA转录通过TFIIIA活性的改变而被下调。

Coordinate regulation of ribosomal component synthesis in Acanthamoeba castellanii: 5S RNA transcription is down regulated during encystment by alteration of TFIIIA activity.

作者信息

Matthews J L, Zwick M G, Paule M R

机构信息

Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins 80523, USA.

出版信息

Mol Cell Biol. 1995 Jun;15(6):3327-35. doi: 10.1128/MCB.15.6.3327.

Abstract

Transcription of large rRNA precursor and 5S RNA were examined during encystment of Acanthamoeba castellanii. Both transcription units are down regulated almost coordinately during this process, though 5S RNA transcription is not as completely shut down as rRNA transcription. The protein components necessary for transcription of 5S RNA and tRNA were determined, and fractions containing transcription factors comparable to TFIIIA, TFIIIB, and TFIIIC, as well as RNA polymerase III and a 3'-end processing activity, were identified. Regulation of 5S RNA transcription could be recapitulated in vitro, and the activities of the required components were compared. In contrast to regulation of precursor rRNA, there is no apparent change during encystment in the activity of the polymerase dedicated to 5S RNA expression. Similarly, the transcriptional and promoter-binding activities of TFIIIC are not altered in parallel with 5S RNA regulation. TFIIIB transcriptional activity is unaltered in encysting cells. In contrast, both the transcriptional and DNA-binding activities of TFIIIA are strongly reduced in nuclear extracts from transcriptionally inactive cells. These results were analyzed in terms of mechanisms for coordinate regulation of rRNA and 5S RNA expression.

摘要

在卡氏棘阿米巴包囊化过程中,对大核糖体RNA前体和5S RNA的转录进行了检测。在这个过程中,两个转录单元几乎是协同下调的,不过5S RNA转录并不像核糖体RNA转录那样完全关闭。确定了5S RNA和tRNA转录所需的蛋白质成分,并鉴定出了含有与TFIIIA、TFIIIB和TFIIIC相当的转录因子的组分,以及RNA聚合酶III和一种3'端加工活性。5S RNA转录的调控可以在体外重现,并对所需组分的活性进行了比较。与前体核糖体RNA的调控不同,在包囊化过程中,负责5S RNA表达的聚合酶的活性没有明显变化。同样,TFIIIC的转录和启动子结合活性也没有随着5S RNA调控而平行改变。在包囊化细胞中,TFIIIB的转录活性没有改变。相比之下,在转录不活跃细胞的核提取物中,TFIIIA的转录和DNA结合活性都大幅降低。根据核糖体RNA和5S RNA表达的协同调控机制对这些结果进行了分析。

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