Neurons modulate gene expression in response to extrinsic signals to enable brain development, cognition, and learning and to process stimuli that regulate systemic physiological functions. This signal-to-gene communication is facilitated by posttranslational modifications such as S-nitrosylation, the covalent attachment of a nitric oxide (NO) moiety to cysteine thiols. In the cerebral cortex, S-nitrosylation of histone deacetylase 2 (HDAC2) is required for gene transcription during neuronal development, but few other nuclear targets of S-nitrosylation have been identified to date. We used S-nitrosothiol resin-assisted capture on NO donor-treated nuclear extracts from rat cortical neurons and identified 614 S-nitrosylated nuclear proteins. Of these, 131 proteins have not previously been shown to be S-nitrosylated in any system, and 555 are previously unidentified targets of S-nitrosylation in neurons. The sites of S-nitrosylation were identified for 59% of the targets, and motifs containing single lysines were found at 33% of these sites. In addition, lysine motifs were necessary for promoting the S-nitrosylation of HDAC2 and methyl-CpG binding protein 3 (MBD3). Moreover, S-nitrosylation of the histone-binding protein RBBP7 was necessary for dendritogenesis of cortical neurons in culture. Together, our findings characterize S-nitrosylated nuclear proteins in neurons and identify S-nitrosylation motifs that may be shared with other targets of NO signaling.

神经元响应外在信号调节基因表达，以促进大脑发育、认知和学习，并处理调节系统生理功能的刺激。这种信号与基因的交流通过翻译后修饰（如 S-亚硝基化）促进了这种信号与基因的交流，一氧化氮 (NO) 部分共价连接到半胱氨酸硫醇上。在大脑皮层中，组蛋白去乙酰化酶 2 (HDAC2) 的 S-亚硝基化是神经元发育过程中基因转录所必需的，但迄今为止，很少有其他的 S-亚硝基化核靶点被发现。我们对大鼠皮质神经元的 NO 供体处理的核提取物使用 S-亚硝基硫醇树脂辅助捕获，并鉴定了 614 个 S-亚硝基化核蛋白。其中，131 种蛋白质以前没有被证明在任何系统中被 S-亚硝基化，和 555 是神经元中 S-亚硝基化的先前未知目标。59% 的目标确定了 S-亚硝基化位点，并且在 33% 的这些位点发现了包含单个赖氨酸的基序。此外，赖氨酸基序是促进 HDAC2 和甲基-CpG 结合蛋白 3 (MBD3) 的 S-亚硝基化所必需的。此外，组蛋白结合蛋白 RBBP7 的 S-亚硝基化对于培养中皮层神经元的树突发生是必要的。总之，我们的研究结果表征了神经元中的 S-亚硝基化核蛋白，并确定了可能与 NO 信号传导的其他目标共享的 S-亚硝基化基序。赖氨酸基序是促进 HDAC2 和甲基-CpG 结合蛋白 3 (MBD3) 的 S-亚硝基化所必需的。此外，组蛋白结合蛋白 RBBP7 的 S-亚硝基化对于培养中皮层神经元的树突发生是必要的。总之，我们的研究结果表征了神经元中的 S-亚硝基化核蛋白，并确定了可能与 NO 信号传导的其他目标共享的 S-亚硝基化基序。赖氨酸基序是促进 HDAC2 和甲基-CpG 结合蛋白 3 (MBD3) 的 S-亚硝基化所必需的。此外，组蛋白结合蛋白 RBBP7 的 S-亚硝基化对于培养中皮层神经元的树突发生是必要的。总之，我们的研究结果表征了神经元中的 S-亚硝基化核蛋白，并确定了可能与 NO 信号传导的其他目标共享的 S-亚硝基化基序。