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大鼠截短重复转录本在非洲爪蟾卵母细胞中的稳定积累。

Stable accumulation of a rat truncated repeat transcript in Xenopus oocytes.

作者信息

Gutierrez-Hartmann A, Baxter J D

出版信息

Proc Natl Acad Sci U S A. 1986 May;83(10):3106-10. doi: 10.1073/pnas.83.10.3106.

Abstract

To define potential mechanisms of expression of middle-repetitive DNA, Xenopus oocytes were employed to examine the rat type 2 and truncated repeat (TR) elements contained in an intron and in the 3'-flanking region of the rat growth hormone gene. These repeats contain significant sequence and structural homology to tRNA genes and, thus, may represent tRNA pseudogenes. Transcripts from the type 2 elements do not accumulate in the cytosol and are found predominantly in the nucleus, whereas those from TR DNA are expressed in the cytosol of neural and pituitary tissues. In HeLa cell extracts, the rat growth hormone type 2 sequences initiate RNA polymerase III transcription resulting in multiple transcripts of 175-970 nucleotides; some of these also contain TR sequences that are present only as downstream structures since the rat growth hormone-TR DNA lacks promoter activity. In Xenopus oocytes the same template also results in multiple transcripts, but with time a single, homogeneous 73-base RNA preferentially accumulates. This RNA probably arises from larger repetitive DNA transcripts as assessed by the kinetics of its formation, its 5' terminus, and the injection of transcripts generated in HeLa cell-free extracts into the oocytes. Sequence analysis of the 73-base RNA suggests that it is a TR transcripts derived from the TR region with tRNA homology. Stable type 2 transcripts were not detected. Thus, type 2 elements are transcribed in the oocytes, but RNAs from them are degraded whereas discrete TR DNA transcripts can be derived from larger RNA molecules and can accumulate in the cytosol due to their preferential stability. These findings indicate that posttranscriptional control mechanisms can operate to direct differential expression of closely related repetitive DNAs and suggest that structures similar to tRNA contained within the TR sequences may allow them to accumulate preferentially in the cytoplasm.

摘要

为了确定中重复DNA表达的潜在机制,采用非洲爪蟾卵母细胞来检测大鼠生长激素基因内含子和3'侧翼区域中包含的大鼠2型和截短重复(TR)元件。这些重复序列与tRNA基因具有显著的序列和结构同源性,因此可能代表tRNA假基因。2型元件的转录本不在细胞质中积累,主要存在于细胞核中,而TR DNA的转录本则在神经和垂体组织的细胞质中表达。在HeLa细胞提取物中,大鼠生长激素2型序列启动RNA聚合酶III转录,产生175 - 970个核苷酸的多个转录本;其中一些还包含仅作为下游结构存在的TR序列,因为大鼠生长激素 - TR DNA缺乏启动子活性。在非洲爪蟾卵母细胞中,相同的模板也会产生多个转录本,但随着时间的推移,一种单一的、均一的73个碱基的RNA优先积累。根据其形成动力学、5'末端以及将HeLa无细胞提取物中产生的转录本注射到卵母细胞中的情况评估,这种RNA可能来自较大的重复DNA转录本。对73个碱基的RNA进行序列分析表明,它是一个来自具有tRNA同源性的TR区域的TR转录本。未检测到稳定的2型转录本。因此,2型元件在卵母细胞中被转录,但其RNA被降解,而离散的TR DNA转录本可以从较大的RNA分子衍生而来,并由于其优先稳定性而在细胞质中积累。这些发现表明,转录后控制机制可以指导密切相关的重复DNA的差异表达,并表明TR序列中包含的类似于tRNA的结构可能使其优先在细胞质中积累。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1fa/323461/6cd714eea53e/pnas00314-0066-a.jpg

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