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对于预mRNA多基化所需的多A) 聚合酶和裂变/特异性因子的分离和表征
Y Takagaki1, L C Ryner, J L Manley
1Department of Biological Sciences, Columbia University, New York, New York 10027.
Cell
|March 11, 1988
概括
研究人员通过分离裂变和多化活动来研究mRNA 3'末端的形成. 恢复这些活动的同时恢复了聚基化中的特异性和效率.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 处理RNA处理RNA处理
背景情况:
- 多基化对于mRNA稳定性和翻译至关重要.
- 聚氨基化mRNA的3'端形成涉及裂变和聚氨基化.
- 已知特定的序列,如AAUAAA,参与了这个过程.
研究的目的:
- 调查控制mRNA 3'末端形成的独特机制和因素.
- 确定裂变和多化活动在特异性和效率方面的作用.
- 要了解这两个过程是如何相互作用的.
主要方法:
- 分解HeLa细胞核提取物以分离酶活性.
- 裂变和多分子聚合酶酶的净化.
- 用于测试各种前体RNA的裂变和多化效率和特异性.
- 复制实验是通过混合分离的酶活动来进行的.
主要成果:
- 分裂和多化是由不同的,可分离的酶活性催化.
- 部分净化的分裂酶保留了AAUAAA信号序列的特异性.
- 分解聚A) 聚合酶失去了特异性.
- 混合裂变和多聚甲基聚合酶分离物恢复了效率和特异性.
- 单独的裂变活性对不同前体的效率有限,但与聚A) 聚合酶结合时会有所改善.
结论:
- 分裂和多化的结合性质对于特定且高效的mRNA 3'端形成至关重要.
- 聚A) 聚合酶在恢复切割效率和特异性方面发挥着作用.
- 可分离的因素有助于复杂的mRNA 3'终端处理过程.
相关概念视频
RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
Nuclear Export of mRNA
Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
mRNA Stability and Gene Expression
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Cis-acting Elements involved in mRNA stability
mRNA Stability and Gene Expression
The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Cis-acting Elements involved in mRNA stability

