NUSAP1 Binds ILF2 to Modulate R-Loop Accumulation and DNA Damage in Prostate Cancer.
NUSAP1 Binds ILF2 to Modulate R-Loop Accumulation and DNA Damage in Prostate Cancer.
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DOI：
10.3390/ijms24076258
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发表时间：
2023-03-26
影响因子：
5.6
通讯作者：
Brooks, James D.
中科院分区：
文献类型：
--
作者：
Chiu, Chun-Lung;Li, Caiyun G.;Verschueren, Erik;Wen, Ru M.;Zhang, Dalin;Gordon, Catherine A.;Zhao, Hongjuan;Giaccia, Amato J.;Brooks, James D.
NUSAP 1的表达增加已被确定为前列腺癌和其他恶性肿瘤的稳健预后生物标志物。我们之前已经证明NUSAP 1受E2 F1的正调控，并促进癌症的侵袭和转移。为了进一步了解NUSAP 1的生物学功能，我们使用亲和纯化和质谱蛋白质组学分析来鉴定NUSAP 1相互作用物。我们在NUSAP 1相互作用组中鉴定了85种独特的蛋白质，包括ILF 2，DHX 9和其他RNA结合蛋白。利用蛋白质组学方法，我们发现NUSAP 1通过与ILF 2的相互作用在维持R环和DNA损伤反应中的功能。共免疫沉淀和共定位共聚焦显微镜验证NUSAP 1与ILF 2和DHX 9，和RNA/DNA杂交的相互作用。我们发现NUSAP 1的微管和带电螺旋结构域是蛋白质-蛋白质相互作用所必需的。单独消耗ILF 2进一步增加喜树碱诱导的R环积累和DNA损伤，NUSAP 1消耗消除了这种作用。在人前列腺癌中，NUSAP 1和ILF 2 mRNA表达水平正相关，升高，并与不良临床结局相关。我们的研究确定了NUSAP 1通过与ILF 2的相互作用在调节R环形成和积累以响应DNA损伤中的新作用，因此提供了潜在的治疗靶点。
Increased expression of NUSAP1 has been identified as a robust prognostic biomarker in prostate cancer and other malignancies. We have previously shown that NUSAP1 is positively regulated by E2F1 and promotes cancer invasion and metastasis. To further understand the biological function of NUSAP1, we used affinity purification and mass spectrometry proteomic analysis to identify NUSAP1 interactors. We identified 85 unique proteins in the NUSAP1 interactome, including ILF2, DHX9, and other RNA-binding proteins. Using proteomic approaches, we uncovered a function for NUSAP1 in maintaining R-loops and in DNA damage response through its interaction with ILF2. Co-immunoprecipitation and colocalization using confocal microscopy verified the interactions of NUSAP1 with ILF2 and DHX9, and RNA/DNA hybrids. We showed that the microtubule and charged helical domains of NUSAP1 were necessary for the protein-protein interactions. Depletion of ILF2 alone further increased camptothecin-induced R-loop accumulation and DNA damage, and NUSAP1 depletion abolished this effect. In human prostate adenocarcinoma, NUSAP1 and ILF2 mRNA expression levels are positively correlated, elevated, and associated with poor clinical outcomes. Our study identifies a novel role for NUSAP1 in regulating R-loop formation and accumulation in response to DNA damage through its interactions with ILF2 and hence provides a potential therapeutic target.
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影响因子：
--
作者：
Gordon CA;Gong X;Ganesh D;Brooks JD
通讯作者：
Brooks JD
影响因子：
2.6
作者：
Haley B;Paunesku T;Protić M;Woloschak GE
通讯作者：
Woloschak GE
影响因子：
64.5
作者：
Cancer Genome Atlas Research Network
通讯作者：
Cancer Genome Atlas Research Network
影响因子：
16
作者：
Brickner, Joshua R.;Garzon, Jada L.;Cimprich, Karlene A.
通讯作者：
Cimprich, Karlene A.
影响因子：
51.1
作者：
Cuzick, Jack;Swanson, Gregory P.;Fisher, Gabrielle;Brothman, Arthur R.;Berney, Daniel M.;Reid, Julia E.;Mesher, David;Speights, V. O.;Stankiewicz, Elzbieta;Foster, Christopher S.;Moller, Henrik;Scardino, Peter;Warren, Jorja D.;Park, Jimmy;Younus, Adib;Flake, Dart D., II;Wagner, Susanne;Gutin, Alexander;Lanchbury, Jerry S.;Stone, Steven
通讯作者：
Stone, Steven