NKAP is a novel RS-related protein that interacts with RNA and RNA binding proteins.
NKAP is a novel RS-related protein that interacts with RNA and RNA binding proteins.
复制标题
DOI：
10.1093/nar/gkt1311
复制
发表时间：
2014-03
影响因子：
14.9
通讯作者：
Noegel AA
中科院分区：
文献类型：
--
作者：
Burgute BD;Peche VS;Steckelberg AL;Glöckner G;Gaßen B;Gehring NH;Noegel AA
NKAP是一种高度保守的蛋白质，在转录抑制、T细胞发育、成熟和获得功能能力以及维持和存活成体造血干细胞中发挥作用。在这里，我们报告了NKAP在剪接中的新作用。使用NKAP特异性抗体，我们发现NKAP定位于核斑点。NKAP在N-末端具有RS基序，随后是高度碱性结构域和C-末端区域的DUF 926结构域。缺失分析表明，基本域是重要的散斑定位。在下拉实验中，我们确定了RNA结合蛋白，RNA解旋酶和剪接因子作为NKAP的相互作用伙伴，其中包括FUS/TLS。FUS/TLS-NKAP相互作用通过NKAP的RS结构域和FUS/TLS的RGG 1和RGG 3结构域发生。我们使用体外剪接测定分析了NKAP与RNA相互作用的能力，发现NKAP结合剪接的信使RNA（mRNA）和未剪接的前mRNA。使用交联和免疫沉淀测序的全基因组分析揭示了NKAP与U1，U4和U 5小核RNA的关联，我们还证明了NKAP的敲低导致前mRNA百分比的增加。我们的研究结果表明NKAP作为核斑点蛋白在RNA剪接和加工中发挥作用。
NKAP is a highly conserved protein with roles in transcriptional repression, T-cell development, maturation and acquisition of functional competency and maintenance and survival of adult hematopoietic stem cells. Here we report the novel role of NKAP in splicing. With NKAP-specific antibodies we found that NKAP localizes to nuclear speckles. NKAP has an RS motif at the N-terminus followed by a highly basic domain and a DUF 926 domain at the C-terminal region. Deletion analysis showed that the basic domain is important for speckle localization. In pull-down experiments, we identified RNA-binding proteins, RNA helicases and splicing factors as interaction partners of NKAP, among them FUS/TLS. The FUS/TLS–NKAP interaction takes place through the RS domain of NKAP and the RGG1 and RGG3 domains of FUS/TLS. We analyzed the ability of NKAP to interact with RNA using in vitro splicing assays and found that NKAP bound both spliced messenger RNA (mRNA) and unspliced pre-mRNA. Genome-wide analysis using crosslinking and immunoprecipitation-seq revealed NKAP association with U1, U4 and U5 small nuclear RNA, and we also demonstrated that knockdown of NKAP led to an increase in pre-mRNA percentage. Our results reveal NKAP as nuclear speckle protein with roles in RNA splicing and processing.
登录
查看更多内容
影响因子：
5.3
作者：
Deckert, Jochen;Hartmuth, Maus;Luehrmann, Reinhard
通讯作者：
Luehrmann, Reinhard
影响因子：
4.5
作者：
Ilagan, Janine O.;Chalkley, Robert J.;Jurica, Melissa S.
通讯作者：
Jurica, Melissa S.
影响因子：
16
作者：
Hegele, Anna;Kamburov, Atanas;Stelzl, Ulrich
通讯作者：
Stelzl, Ulrich
DOI：
10.1016/j.bbrc.2003.09.074
发表时间：
2003-10-24
影响因子：
3.1
作者：
Chen, DY;Li, ZQ;Shu, HB
通讯作者：
Shu, HB
影响因子：
4.5
作者：
Bessonov, Sergey;Anokhina, Maria;Luehrmann, Reinhard
通讯作者：
Luehrmann, Reinhard