100千道尔顿的U5小核核糖核蛋白（hPrp28p）通过其ATP酶位点与5'剪接位点接触。
The 100-kda U5 snRNP protein (hPrp28p) contacts the 5' splice site through its ATPase site.
作者信息
Ismaïli N, Sha M, Gustafson E H, Konarska M M
机构信息
The Rockefeller University, New York, New York 10021, USA.
出版信息
RNA. 2001 Feb;7(2):182-93. doi: 10.1017/s1355838201001807.
Abstract
To identify splicing factors in proximity of the 5' splice site (5'SS), we followed a crosslinking profile of site-specifically modified, photoreactive RNA substrates. Upon U4/U5/U6 snRNP addition, the 5'SS RNA crosslinks in an ATP-dependent manner to U6 snRNA, an unidentified protein p27, and the 100-kDa U5 snRNP protein, a human ortholog of an ATPase/RNA helicase yPrp28p. The 5'SS:hPrp28p crosslink maps to the highly conserved TAT motif in proximity of the ATP-binding site in hPrp28p. We propose that hPrp28p acts as a helicase to unwind the 5'SS:U1 snRNA duplex, and at the same time as a 5'SS translocase, which, upon NTP-dependent conformational change, positions the 5'SS for pairing with U6 snRNA within the spliceosome. This repositioning of the 5'SS takes place regardless of whether the 5'SS is originally duplexed with U1 snRNA.
摘要
为了鉴定靠近5'剪接位点（5'SS）的剪接因子，我们追踪了位点特异性修饰的光反应性RNA底物的交联图谱。添加U4/U5/U6 snRNP后，5'SS RNA以ATP依赖的方式与U6 snRNA、一种未鉴定的蛋白质p27以及100 kDa的U5 snRNP蛋白质交联，该蛋白质是ATP酶/RNA解旋酶yPrp28p的人类同源物。5'SS:hPrp28p交联定位于hPrp28p中靠近ATP结合位点的高度保守的TAT基序。我们提出，hPrp28p作为解旋酶解开5'SS:U1 snRNA双链体，同时作为5'SS转位酶，在NTP依赖的构象变化后，将5'SS定位以便与剪接体内的U6 snRNA配对。无论5'SS最初是否与U1 snRNA双链化，5'SS的这种重新定位都会发生。
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