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9E3 mRNA在正常细胞和劳氏肉瘤病毒转化细胞中的加工及其稳定性调控。

Processing of 9E3 mRNA and regulation of its stability in normal and Rous sarcoma virus-transformed cells.

作者信息

Stoeckle M Y, Hanafusa H

机构信息

Laboratory of Molecular Oncology, Rockefeller University, New York, New York 10021.

出版信息

Mol Cell Biol. 1989 Nov;9(11):4738-45. doi: 10.1128/mcb.9.11.4738-4745.1989.

DOI:10.1128/mcb.9.11.4738-4745.1989
PMID:2557539
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC363621/
Abstract

We studied the expression of 9E3 mRNA, which is known to be induced in chicken embryo fibroblasts by p60v-src activity and by serum. In addition to full-length 9E3 mRNA, we identified several smaller RNAs that hybridized with 9E3 cDNA. One of these RNAs hybridized with a 5' 9E3 cDNA probe but not with a 3' cDNA probe. The other hybridized with a 3' cDNA probe but lacked 5' sequences, including the entire 9E3 coding region. Only the latter RNA was polyadenylylated, as determined by RNase H digestion in the presence of oligo(dT). The level of the small RNAs increased after treatment with cycloheximide and actinomycin D, indicating that the small RNAs were produced by processing of preexisting transcripts. The derivation of the small RNAs from 9E3 mRNA rather than from a related gene was confirmed by S1 nuclease analysis. The 3' terminus of the 5' RNA and the 5' terminus of the 3' RNA mapped to the same position, which suggested that the small RNAs were formed by endonucleolytic cleavage of 9E3 mRNA at a specific site in the 3' noncoding region. We also found that the stability of 9E3 mRNA was increased after serum stimulation and was greater in Rous sarcoma virus-transformed than in uninfected cells. The relative amount of the small RNAs as compared with the full-length transcript was greatest under conditions in which the full-length transcript was least stable. These data suggest that site-specific endonucleolytic cleavage regulates the stability of 9E3 mRNA.

摘要

我们研究了9E3 mRNA的表达，已知其在鸡胚成纤维细胞中可由p60v-src活性和血清诱导产生。除了全长9E3 mRNA外，我们还鉴定出了几种与9E3 cDNA杂交的较小RNA。其中一种RNA与5' 9E3 cDNA探针杂交，但不与3' cDNA探针杂交。另一种则与3' cDNA探针杂交，但缺乏5'序列，包括整个9E3编码区。通过在寡聚(dT)存在下进行RNase H消化确定，只有后一种RNA是聚腺苷酸化的。用环己酰亚胺和放线菌素D处理后，小RNA的水平升高，表明这些小RNA是由预先存在的转录本加工产生的。通过S1核酸酶分析证实了小RNA来源于9E3 mRNA而非相关基因。5' RNA的3'末端和3' RNA的5'末端定位于同一位置，这表明小RNA是由9E3 mRNA在3'非编码区的特定位点进行内切核酸酶切割形成的。我们还发现，血清刺激后9E3 mRNA的稳定性增加，并且在劳氏肉瘤病毒转化的细胞中比未感染细胞中的稳定性更高。与全长转录本相比，小RNA的相对量在全长转录本最不稳定的条件下最大。这些数据表明，位点特异性内切核酸酶切割调节9E3 mRNA的稳定性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/c5ca87abc823/molcellb00059-0167-c.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/99b7b2c8b615/molcellb00059-0167-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/7854a9e43aea/molcellb00059-0167-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/c5ca87abc823/molcellb00059-0167-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/c8b8956be9ca/molcellb00059-0164-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/30e1616ee638/molcellb00059-0164-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/304e229b11bc/molcellb00059-0164-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/3226afae3df1/molcellb00059-0165-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/be66d10924e4/molcellb00059-0165-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/c35f2991e922/molcellb00059-0166-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/9c877cd60e25/molcellb00059-0166-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/99b7b2c8b615/molcellb00059-0167-a.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a1e/363621/c5ca87abc823/molcellb00059-0167-c.jpg

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