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小鼠类病毒30S(VL30)RNA的进一步特性研究:逆转录的起始与细胞内合成

Further characterization of virus-like 30S (VL30) RNA of mice: initiation of reverse transcription and intracellular synthesis.

作者信息

Dolberg D, Fan H

出版信息

J Gen Virol. 1981 Jun;54(Pt 2):281-91. doi: 10.1099/0022-1317-54-2-281.

Abstract

We have studied the virus-like 30S (VL30) RNA sequences of mice. Previous work has shown that these sequences are coded in the mouse genome, expressed in some normal cells and released as pseudotypic particles from cells producing murine C-type retroviruses. VL30 sequences have some similarities to standard retrovirus RNA, but differences also exist. To further assess the similarities and differences, several aspects of VL30-specific metabolism were investigated. We studied the initiation of VL30-specific DNA synthesis during an endogenous reverse transcriptase reaction. Short initial VL30-specific cDNA transcripts were covalently attached to RNA as measured by equilibrium banding in caesium sulphate density gradients. Therefore, reverse transcription of VL30-specific cDNA is initiated by an RNA primer. The intracellular synthesis of VL30 RNA was investigated by pulse labelling uninfected JLS-V9 cells with 3H-uridine. Hybridization of the pulse-labelled nuclear RNA indicated that the major VL30-specific RNA evident after a 15 min label was the same size as the mature VL30 RNA. Thus, VL30 RNA is apparently not synthesized via a higher mol. wt. precursor. Both of these results demonstrate similarity of VL30 RNA sequences to standard retroviruses. One unique feature of VL30 RNA was detected. JLS-V9 cells contained both the monomeric VL30 RNA and a hydrogen-bonded 38S form which yielded the monomer when denatured. This contrasts with standard murine leukaemia virus which is only found as a monomer within cells.

摘要

我们研究了小鼠的病毒样30S(VL30)RNA序列。先前的研究表明,这些序列编码于小鼠基因组中,在一些正常细胞中表达,并作为假型颗粒从产生鼠C型逆转录病毒的细胞中释放出来。VL30序列与标准逆转录病毒RNA有一些相似之处,但也存在差异。为了进一步评估这些异同,我们研究了VL30特异性代谢的几个方面。我们研究了内源性逆转录酶反应过程中VL30特异性DNA合成的起始情况。通过硫酸铯密度梯度平衡沉降法测定,最初的短VL30特异性cDNA转录本与RNA共价连接。因此,VL30特异性cDNA的逆转录是由RNA引物起始的。通过用3H-尿苷脉冲标记未感染的JLS-V9细胞来研究VL30 RNA的细胞内合成。脉冲标记的核RNA杂交表明,标记15分钟后明显的主要VL30特异性RNA与成熟VL30 RNA大小相同。因此,VL30 RNA显然不是通过更高分子量的前体合成的。这两个结果都证明了VL30 RNA序列与标准逆转录病毒的相似性。我们还检测到了VL30 RNA的一个独特特征。JLS-V9细胞同时含有单体VL30 RNA和一种氢键结合的38S形式,变性后可产生单体。这与标准鼠白血病病毒不同,后者在细胞内仅以单体形式存在。

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