The C-terminal region of hPrp8 interacts with the conserved GU dinucleotide at the 5' splice site.
The C-terminal region of hPrp8 interacts with the conserved GU dinucleotide at the 5' splice site.
复制标题
hPrp8 的 C 末端区域与 5 剪接位点的保守 GU 二核苷酸相互作用。
DOI：
10.1017/s1355838299981785
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Konarska,MM
中科院分区：
文献类型：
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作者：
Reyes,JL;Gustafson,EH;Luo,HR;Moore,MJ;Konarska,MM
U5 snRNP蛋白hPrp8与剪接复合体B中的5‘剪接位点（5’ ss） RNA在反式和顺式剪接反应中形成uv诱导的交联。酵母和人类Prp8在剪接过程中都与5'SS、分支位点、聚嘧啶通道和3'SS相互作用。为了开始定义Prp8的功能域，我们绘制了hPrp8蛋白内5'SS交联的位点。免疫沉淀分析将交联位点限制在hPrp8的c端5060-kDa片段上。此外，将不同蛋白水解试剂生成的含交联肽与hPrp8序列预测的片段模式进行大小比较，可以将交联映射到hPrp8 c端部分（位置1894-1898）的五个氨基酸。5'SS:hPrp8交联位点位于先前定义的yPrp8多聚嘧啶束识别区域内，这表明Prp8的重叠区域可能参与了5'SS和多聚嘧啶束识别事件。在其他已知的Prp8相互作用的背景下，这些结果表明该蛋白可能参与剪接体催化中心的形成。
A U5 snRNP protein, hPrp8, forms a UV-induced crosslink with the 5' splice site (5'SS) RNA within splicing complex B assembled in trans- as well as in cis-splicing reactions. Both yeast and human Prp8 interact with the 5'SS, branch site, polypyrimidine tract, and 3'SS during splicing. To begin to define functional domains in Prp8 we have mapped the site of the 5'SS crosslink within the hPrp8 protein. Immunoprecipitation analysis limited the site of crosslink to the C-terminal 5060-kDa segment of hPrp8. In addition, size comparison of the crosslink-containing peptides generated with different proteolytic reagents with the pattern of fragments predicted from the hPrp8 sequence allowed for mapping of the crosslink to a stretch of five amino acids in the C-terminal portion of hPrp8 (positions 1894-1898). The site of the 5'SS:hPrp8 crosslink falls within a segment spanning the previously defined polypyrimidine tract recognition domain in yPrp8, suggesting that an overlapping region of Prp8 may be involved both in the 5'SS and polypyrimidine tract recognition events. In the context of other known interactions of Prp8, these results suggest that this protein may participate in formation of the catalytic center of the spliceosome.