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多能胚胎癌细胞系中核和多核糖体聚腺苷酸化RNA的复杂性

Complexity of nuclear and polysomal polyadenylated RNA in a pluripotent embryonal carcinoma cell line.

作者信息

Jacquet M, Affara N A, Robert B, Jakob H, Jacob F, Gros F

出版信息

Biochemistry. 1978 Jan 10;17(1):69-79. doi: 10.1021/bi00594a010.

Abstract

The base-sequence complexities and relative abundance of polysomal and nuclear polyadenylated [poly(A+)] RNA sequences have been analyzed in a pluripotent embryonal carcinoma cell line. Polysomal RNA and nuclear poly(A+) RNA have a complexity representing respectively 0.5% and 2.5% of the single copy component of haploid mouse DNA (1.8 X 10(6) K base pairs). By hybridization with specific cDNAs, three abundance classes were found in polysomal poly(A+) RNA, representing respectively 31%, 33%, and 36% of the RNA, with base sequence complexities of 0.1 X 10(3), 0.9 X 10(3), and 14.5 X 10(3) kilobases. This corresponds to 7000-8000 different mRNA species of an average length of 2000 nucleotides, present on an average of 5 to 600 copies per cell. In nuclear RNA, a major class of abundance was found with a complexity of 100 X 10(3) kilobases, each sequence being present in 1 copy per nucleus. The majority of the polysomal poly(A+) RNA sequences are represented in the nuclear poly(A+) RNA but are present in a more restricted range of relative abundance implying posttranscriptional mechanisms of quantitative modulation: polysomal RNA sequences appear to be preferentially transcribed into nuclear cDNA suggesting a preferential location of these sequences close to poly(A) sequences. The presence of a specialized gene product, globin specific RNA, could not be detected either in the nuclear or polysomal compartments of embryonal carcinoma cells, even at levels that would have detected one sequence per 50 cells.

摘要

在一个多能胚胎癌细胞系中,对多聚核糖体和细胞核多聚腺苷酸化[poly(A+)]RNA序列的碱基序列复杂性和相对丰度进行了分析。多聚核糖体RNA和细胞核poly(A+)RNA的复杂性分别相当于单倍体小鼠DNA单拷贝组分(1.8×10(6)千碱基对)的0.5%和2.5%。通过与特异性cDNA杂交,在多聚核糖体poly(A+)RNA中发现了三个丰度类别,分别占RNA的31%、33%和36%,碱基序列复杂性分别为0.1×10(3)、0.9×10(3)和14.5×10(3)千碱基。这相当于7000 - 8000种不同的mRNA种类,平均长度为2000个核苷酸,每个细胞平均存在5至600个拷贝。在细胞核RNA中,发现了一个主要的丰度类别,复杂性为100×10(3)千碱基,每个序列在每个细胞核中存在1个拷贝。大多数多聚核糖体poly(A+)RNA序列在细胞核poly(A+)RNA中也有,但相对丰度范围更窄,这意味着存在转录后定量调节机制:多聚核糖体RNA序列似乎优先转录到细胞核cDNA中,表明这些序列优先位于靠近poly(A)序列的位置。即使在每50个细胞中能检测到一个序列的水平,也未在胚胎癌细胞的细胞核或多聚核糖体组分中检测到一种特殊的基因产物——珠蛋白特异性RNA。

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