Multiple cis elements regulate an alternative splicing event at 4.1R pre-mRNA during erythroid differentiation
Multiple cis elements regulate an alternative splicing event at 4.1R pre-mRNA during erythroid differentiation
复制标题
DOI：
10.1182/blood.v98.13.3809
复制
发表时间：
2001-12-15
期刊：
影响因子：
20.3
通讯作者：
Baklouti, F
中科院分区：
文献类型：
--
作者：
Deguillien, M;Huang, SC;Baklouti, F
成熟蛋白 4.1 R 信使 RNA (mRNA) 中外显子 16 的包含是红细胞膜生物发生中的关键事件。它发生在红细胞发育后期，并导致包含稳定血影蛋白/肌动蛋白晶格所需的 10-kd 结构域。在这项研究中，在小鼠红白血病细胞中建立了一个实验模型，该模型可重现转染小基因的内源性外显子 16 剪接模式。外显子 16 在预分化时被排除，而在诱导后主要被包含。这表明小基因包含足以进行剪接调节的外显子和邻接内含子序列。对位于外显子和侧翼内含子内的 cls 作用调控序列进行了系统分析。结果表明：（1）4.1R前体mRNA的上游内含子是外显子识别所必需的，它显示2个增强子元件，一个作用于分化细胞的远端元件和一个位于受体位点之前25个核苷酸内的近端组成型增强子； (2)外显子本身含有强的本构剪接沉默子； (3)外显子有较弱的5'剪接位点； (4)下游内含子含有至少2个作用于分化细胞的剪接增强子元件，一个位于5'剪接位点附近的近端元件，以及一个含有3个拷贝的UGCAUG基序的远端元件。这些结果表明，阴性和阳性元件之间的相互作用可能决定外显子 16 的包含或排除。晚期红细胞分化中增强子元件的激活可能在外显子 16 的保留中发挥重要作用。(C) 2001，美国血液学会。
The inclusion of exon 16 in the mature protein 4.1 R messenger RNA (mRNA) is a critical event in red blood cell membrane biogenesis. It occurs during late erythroid development and results in inclusion of the 10-kd domain needed for stabilization of the spectrin/actin lattice. In this study, an experimental model was established in murine erythroleukemia cells that reproduces the endogenous exon 16 splicing patterns from a transfected minigene. Exon 16 was excluded in predifferentiated and predominantly included after induction. This suggests that the minigene contained exon and abutting intronic sequences sufficient for splicing regulation. A systematic analysis of the cls-acting regulatory sequences that reside within the exon and flanking introns was performed. Results showed that (1) the upstream intron of 4.1 R pre-mRNA is required for exon recognition and it displays 2 enhancer elements, a distal element acting in differentiating cells and a proximal constitutive enhancer that resides within the 25 nucleotides preceding the acceptor site; (2) the exon itself contains a strong constitutive splicing silencer; (3) the exon has a weak 5' splice site; and (4) the downstream intron contains at least 2 splicing enhancer elements acting in differentiating cells, a proximal element at the vicinity of the 5' splice site, and a distal element containing 3 copies of the UGCAUG motif. These results suggest that the interplay between negative and positive elements may determine the inclusion or exclusion of exon 16. The activation of the enhancer elements in late erythroid differentiation may play an important role in the retention of exon 16. (C) 2001 by The American Society of Hematology.