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营养期粗壮真核四膜虫细胞中大核基因的转录速率和转录本稳定性

Transcription rates and transcript stabilities of macronuclear genes in vegetative Euplotes crassus cells.

作者信息

Brandt A, Klein A

机构信息

Department of Biology, Phillipps-University, Marburg, Germany.

出版信息

J Eukaryot Microbiol. 1995 Nov-Dec;42(6):691-6. doi: 10.1111/j.1550-7408.1995.tb01617.x.

Abstract

In hypotrichous ciliates such as Euplotes crassus genes in the transcriptionally active macronucleus are present on individual minichromosomes which occur in gene-specific copy numbers. This different degree of gene amplification can be understood as a means to preset the expression potential of the respective genetic information. In addition, the actual steady state transcript amounts are governed by the transcription rates and transcript stabilities. To establish the relative effects of these three parameters the copy numbers of genes transcribed by the three different polymerases were determined. The transcript levels of growing or starving vegetative cells were then determined, and nuclear run-on assays were performed to determine the transcription rates of the genes in the different nutritional states. A weak correlation between the gene copy numbers and transcription rates was found. The transcripts of genes synthesized by RNA polymerase II exhibited different stabilities upon starvation of the cells, compared to the supposedly stable ribosomal 5S and 26S RNA. Refeeding of the cells after starvation also resulted in a differential response with respect to the accumulation of the transcripts of different genes transcribed by RNA polymerase II, which can be interpreted in the context of the gene functions.

摘要

在诸如粗壮真核游仆虫这样的稀毛纲纤毛虫中,转录活跃的大核中的基因存在于个别微型染色体上,这些微型染色体以基因特异性拷贝数出现。这种不同程度的基因扩增可被理解为一种预设各自遗传信息表达潜力的方式。此外,实际的稳态转录本量由转录速率和转录本稳定性决定。为了确定这三个参数的相对影响,测定了由三种不同聚合酶转录的基因的拷贝数。然后测定了生长或饥饿的营养细胞的转录水平,并进行了细胞核连续标记分析以确定不同营养状态下基因的转录速率。发现基因拷贝数与转录速率之间存在微弱的相关性。与假定稳定的核糖体5S和26S RNA相比,细胞饥饿时由RNA聚合酶II合成的基因的转录本表现出不同的稳定性。饥饿后细胞重新喂食也导致了RNA聚合酶II转录的不同基因转录本积累方面的差异反应,这可以在基因功能的背景下进行解释。

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