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猿猴病毒40小t抗原前体mRNA的有效剪接需要多个顺式作用序列元件。

Multiple cis-acting sequence elements are required for efficient splicing of simian virus 40 small-t antigen pre-mRNA.

作者信息

Fu X Y, Colgan J D, Manley J L

机构信息

Department of Biological Sciences, Columbia University, New York, New York 10027.

出版信息

Mol Cell Biol. 1988 Sep;8(9):3582-90. doi: 10.1128/mcb.8.9.3582-3590.1988.

DOI:10.1128/mcb.8.9.3582-3590.1988
PMID:2851720
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC365413/
Abstract

We have determined the effects of a number of mutations in the small-t antigen mRNA intron on the alternative splicing pattern of the simian virus 40 early transcript. Expansion of the distance separating the small-t pre-mRNA lariat branch point and the shared large T-small t 3' splice site from 18 to 29 nucleotides (nt) resulted in a relative enhancement of small-t splicing in vivo. This finding, coupled with the observation that large-T pre-RNA splicing in vitro was not affected by this expansion, suggests that small-t splicing is specifically constrained by a short branch point-3' splice site distance. Similarly, the distance separating the 5' splice site and branch point (48 nt) was found to be at or near a minimum for small-t splicing, because deletions in this region as small as 2 nt dramatically reduced the ratio of small-t to large-T mRNA that accumulated in transfected cells. Finally, a specific sequence within the small-t intron, encompassing the upstream branch sites used in large-T splicing, was found to be an important element in the cell-specific pattern of early alternative splicing. Substitutions within this region reduced the ratio of small-t to large-T mRNA produced in HeLa cells but had only minor effects in human 293 cells.

摘要

我们已经确定了猴病毒40早期转录本的小t抗原mRNA内含子中的一些突变对可变剪接模式的影响。将小t前体mRNA套索状分支点与共享的大T-小t 3'剪接位点之间的距离从18个核苷酸（nt）扩展到29个核苷酸，导致体内小t剪接相对增强。这一发现，再加上体外大T前体RNA剪接不受此扩展影响的观察结果，表明小t剪接受到短分支点-3'剪接位点距离的特异性限制。同样，发现5'剪接位点与分支点之间的距离（48 nt）对于小t剪接处于或接近最小值，因为该区域小至2 nt的缺失会显著降低转染细胞中积累的小t与大T mRNA的比例。最后，发现小t内含子内包含大T剪接中使用的上游分支位点的特定序列是早期可变剪接细胞特异性模式中的一个重要元件。该区域内的替代降低了HeLa细胞中产生的小t与大T mRNA的比例，但对人293细胞只有轻微影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/96b5bb7abcf2/molcellb00069-0028-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/200af96f1a68/molcellb00069-0025-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/41d19d80d347/molcellb00069-0026-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/1a64d14fd562/molcellb00069-0027-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/6a353537e3b8/molcellb00069-0027-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/96b5bb7abcf2/molcellb00069-0028-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/200af96f1a68/molcellb00069-0025-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/41d19d80d347/molcellb00069-0026-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/1a64d14fd562/molcellb00069-0027-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/6a353537e3b8/molcellb00069-0027-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52a6/365413/96b5bb7abcf2/molcellb00069-0028-a.jpg

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