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未感染鸡细胞中内源性病毒基因RNA转录本的遗传变异。

Genetic variation in the RNA transcripts of endogenous virus genes in uninfected chicken cells.

作者信息

Wang S Y, Hayward W S, Hanafusa H

出版信息

J Virol. 1977 Oct;24(1):64-73. doi: 10.1128/JVI.24.1.64-73.1977.

Abstract

Uninfected cells from two different phenotypes of chicken embryos express significant amounts of endogenous viral information, though they do not produce virus particles. Cells of the phenotype gs(+)chf(+) are positive for both group-specific (gs) antigens and chicken helper factor (chf) activity, whereas cells of a second phenotype, gs(L)chf(+)(h(E)), demonstrate noncoordinate expression of these two viral activities (very low amounts of gs antigens, but extremely high helper activity). RNA from these cells was analyzed to determine the size, genetic content, and relative abundance of virus-specific RNAs in cells of each phenotype. Two major size classes of polyadenylic acid-containing RNA, homologous to the avian leukosis virus genome, were detectable in cells of both types. The larger RNA, which contained most of the sequences of the leukosis virus genome, was of different sizes in the two phenotypes, 31S in gs(+)chf(+) cells but 35S in the noncoordinate cell type. Analysis of the viral RNA with gene-specific complementary DNA probes revealed the following characteristics. (i) The 31S RNA appeared to lack portions of the gag and pol genes. (ii) A smaller RNA species, which sedimented at 21S in both cell types, was a transcript of the 3'-proximal portion of the viral genome, consisting of the env gene and the "common" sequences. (iii) The amount of env-specific RNA in the 21S region was more than six times higher in the noncoordinate cell type than in the gs(+)chf(+) cells; this difference was concordant with the 5- to 10-fold higher chf activity in the noncoordinate cells. (iv) The endogenous viral RNA in uninfected cells and the RNA from Rous-associated virus-0 virions hybridized only partially with DNA complementary to the common region of the Rous-associated virus-2 genome, whereas the RNA of all exogenous viruses tested hybridized almost completely to this complementary DNA. Small amounts of src-specific polyadenylated RNA were also present in uninfected chicken cells. This RNA sedimented as a single peak at 26S and was not covalently linked to any other identifiable virus-specific RNA sequences. The amount of src RNA was the same in the above two types of expression-positive cells and also in cells that were gs(-)chf(-), indicating that the transcription of the cellular sequences homologous to the src gene is independent of the transcription of the other endogenous viral genes.

摘要

来自两种不同表型鸡胚的未感染细胞表达大量内源性病毒信息,尽管它们不产生病毒颗粒。gs(+)chf(+)表型的细胞对群特异性(gs)抗原和鸡辅助因子(chf)活性均呈阳性,而第二种表型gs(L)chf(+)(h(E))的细胞则表现出这两种病毒活性的非协调性表达(gs抗原量非常低,但辅助活性极高)。对这些细胞的RNA进行分析,以确定每种表型细胞中病毒特异性RNA的大小、遗传内容和相对丰度。在两种类型的细胞中均检测到与禽白血病病毒基因组同源的两类主要含多聚腺苷酸的RNA。较大的RNA包含白血病病毒基因组的大部分序列,在两种表型中的大小不同,在gs(+)chf(+)细胞中为31S,而在非协调性细胞类型中为35S。用基因特异性互补DNA探针分析病毒RNA揭示了以下特征。(i)31S RNA似乎缺少gag和pol基因的部分。(ii)一种较小的RNA种类,在两种细胞类型中均沉降在21S,是病毒基因组3'近端部分的转录本,由env基因和“共同”序列组成。(iii)21S区域中env特异性RNA的量在非协调性细胞类型中比在gs(+)chf(+)细胞中高六倍以上;这种差异与非协调性细胞中高5至10倍的chf活性一致。(iv)未感染细胞中的内源性病毒RNA和来自劳斯相关病毒0病毒粒子的RNA仅与与劳斯相关病毒2基因组共同区域互补的DNA部分杂交,而所有测试的外源性病毒的RNA几乎完全与这种互补DNA杂交。未感染的鸡细胞中也存在少量src特异性多聚腺苷酸化RNA。这种RNA在26S处沉降为单峰,并且不与任何其他可识别的病毒特异性RNA序列共价连接。上述两种表达阳性细胞以及gs(-)chf(-)细胞中的src RNA量相同,这表明与src基因同源的细胞序列的转录独立于其他内源性病毒基因的转录。

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