Human RNPS1 and its associated factors: A versatile alternative pre-mRNA splicing regulator in vivo
Human RNPS1 and its associated factors: A versatile alternative pre-mRNA splicing regulator in vivo
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DOI：
10.1128/mcb.24.3.1174-1187.2004
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发表时间：
2004-02-01
影响因子：
5.3
通讯作者：
Mayeda, A
中科院分区：
文献类型：
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作者：
Sakashita, E;Tatsumi, S;Mayeda, A
人类RNPS1最初被纯化并被表征为pre-mRNA剪接激活因子，其在剪接后过程中的作用最近也被提出。为了寻找与RNPS1功能相互作用的因子，我们使用人类cDNA文库进行了酵母双杂交筛选。确定了四个因子：p54（也称为SRp54， SR蛋白家族的成员），人类变压器2beta （hTra2beta，一种外显子剪接增强子结合蛋白），hLucA （U1 snRNP的潜在成分）和pinin（也称为DRS和MemA，一种定位于核斑点的蛋白）。RNPS1含有富含丝氨酸(S)结构域的n端区域、中央RNA识别基序（RRM）和c端富含精氨酸/丝氨酸/脯氨酸（RS/P）结构域分别与p54、pinin和hTra2beta相互作用。在体外和体内验证了RNPS1与这些因子之间的蛋白结合。HeLa细胞中RNPS1的过表达诱导了模型β -珠蛋白前mrna和人tra-2 β前mrna的外显子跳变。RNPS1与p54的共表达协同刺激ATP合成酶γ -亚基前mrna的外显子包涵。RS/P结构域和RRM对于外显子跳跃活性是必需的，而S结构域对于与p54的协同作用是重要的。RNPS1似乎是一个多功能因子，调节各种前mrna的选择性剪接。
Human RNPS1 was originally purified and characterized as a pre-mRNA splicing activator, and its role in the postsplicing process has also been proposed recently. To search for factors that functionally interact with RNPS1, we performed a yeast two-hybrid screen with a human cDNA library. Four factors were identified: p54 (also called SRp54; a member of the SR protein family), human transformer 2beta (hTra2beta; an exonic splicing enhancer-binding protein), hLucA (a potential component of U1 snRNP), and pinin (also called DRS and MemA; a protein localized in nuclear speckles). The N-terminal region containing the serine-rich (S) domain, the central RNA recognition motif (RRM), and the C-terminal arginine/serine/proline-rich (RS/P) domain of RNPS1 interact with p54, pinin, and hTra2beta, respectively. Protein-protein binding between RNPS1 and these factors was verified in vitro and in vivo. Overexpression of RNPS1 in HeLa cells induced exon skipping in a model beta-globin pre-mRNA and a human tra-2beta pre-mRNA. Coexpression of RNPS1 with p54 cooperatively stimulated exon inclusion in an ATP synthase gamma-subunit pre-mRNA. The RS/P domain and RRM are necessary for the exon-skipping activity, whereas the S domain is important for the cooperative effect with p54. RNPS1 appears to be a versatile factor that regulates alternative splicing of a variety of pre-mRNAs.