INTERACTIONS OF SMALL NUCLEAR RNAS WITH PRECURSOR MESSENGER-RNA DURING INVITRO SPLICING
INTERACTIONS OF SMALL NUCLEAR RNAS WITH PRECURSOR MESSENGER-RNA DURING INVITRO SPLICING
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DOI：
10.1126/science.1411506
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发表时间：
1992-09-25
期刊：
影响因子：
56.9
通讯作者：
STEITZ, JA
中科院分区：
文献类型：
--
作者：
WASSARMAN, DA;STEITZ, JA
前体信使RNA剪接需要多种因子，包括U1、U2、U4、U 5和U6小核RNA。使用交联试剂补骨脂素来分析这些RNA与HeLa核提取物中腺病毒前体信使RNA的相互作用。内源性U2-U4-U6可交联复合物在与前体信使RNA孵育后解离。在剪接过程中，U1、U2、U 5和U6与前体信使RNA交联，U2、U 5和U6与切除的内含子交联。U2还与U6和前体信使RNA形成双重交联复合物。U1、U 5和U6与前体信使RNA的交联映射到5'剪接位点附近的内含子序列，而U2交联映射到分支位点。交联形成和消失的动力学描绘了剪接体中小RNA作用的时间途径。小核RNA和前体信使RNA中的保守序列在交联位点处的潜在碱基配对相互作用表明，5'剪接位点在第一次切割事件之前的几个步骤中被限定。
Precursor messenger RNA splicing requires multiple factors including U1, U2, U4, U5, and U6 small nuclear RNA's. The crosslinking reagent psoralen was used to analyze the interactions of these RNA's with an adenovirus precursor messenger RNA in HeLa nuclear extract. An endogenous U2-U4-U6 crosslinkable complex dissociated upon incubation with precursor messenger RNA. During splicing, U1, U2, U5, and U6 became crosslinked to precursor messenger RNA and U2, U5, and U6 became crosslinked to excised lariat intron. U2 also formed a doubly crosslinked complex with U6 and precursor messenger RNA. The U1, U5, and U6 crosslinks to the precursor messenger RNA mapped to intron sequences near the 5' splice site, whereas the U2 crosslink mapped to the branch site. The kinetics of crosslink formation and disappearance delineates a temporal pathway for the action of small RNA's in the spliceosome. Potential base pairing interactions between conserved sequences in the small nuclear RNA's and precursor messenger RNA at the sites of crosslinking suggest that the 5' splice site is defined in several steps prior to the first cleavage event.