Cockayne Syndrome is a segmental premature aging syndrome, which can be caused by loss of function of the CSB protein. CSB is essential for genome maintenance and has numerous interaction partners with established roles in different DNA repair pathways including transcription coupled nucleotide excision repair and base excision repair. Here, we describe a new interaction partner for CSB, the DNA glycosylase NEIL2. Using both cell extracts and recombinant proteins, CSB and NEIL2 were found to physically interact independently of DNA. We further found that CSB is able to stimulate NEIL2 glycosylase activity on a 5-hydroxyl uracil lesion in a DNA bubble structure substrate in vitro. A novel 4,6-diamino-5-formamidopyrimidine (FapyA) specific incision activity of NEIL2 was also stimulated by CSB. To further elucidate the biological role of the interaction, immunofluorescence studies were performed, showing an increase in cytoplasmic CSB and NEIL2 co-localization after oxidative stress. Additionally, stalling of the progression of the transcription bubble with α-amanitin resulted in increased co-localization of CSB and NEIL2. Finally, CSB knockdown resulted in reduced incision of 8-hydroxyguanine in a DNA bubble structure using whole cell extracts. Taken together, our data supports a biological role for CSB and NEIL2 in transcription associated base excision repair.

中文翻译：

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Cockayne 综合征 B 组蛋白刺激 NEIL2 DNA 糖基化酶活性。

Cockayne 综合征是一种节段性过早衰老综合征，可由 CSB 蛋白功能丧失引起。CSB 对基因组维护至关重要，并且具有许多相互作用伙伴，它们在不同的 DNA 修复途径中发挥着既定作用，包括转录偶联核苷酸切除修复和碱基切除修复。在这里，我们描述了 CSB，DNA 糖基化酶 NEIL2 的新相互作用伙伴。使用细胞提取物和重组蛋白，发现 CSB 和 NEIL2 独立于 DNA 进行物理相互作用。我们进一步发现 CSB 能够在体外刺激 DNA 气泡结构底物中 5-羟基尿嘧啶损伤的 NEIL2 糖基化酶活性。CSB 还刺激了 NEIL2 的新型 4,6-二氨基-5-甲酰胺嘧啶 (FapyA) 特异性切口活性。为了进一步阐明相互作用的生物学作用，进行了免疫荧光研究，显示氧化应激后细胞质 CSB 和 NEIL2 共定位增加。此外，用α-鹅膏菌素阻止转录气泡的进展导致CSB和NEIL2的共定位增加。最后，使用全细胞提取物，CSB 敲低导致 DNA 气泡结构中 8-羟基鸟嘌呤的切口减少。总之，我们的数据支持 CSB 和 NEIL2 在转录相关碱基切除修复中的生物学作用。最后，使用全细胞提取物，CSB 敲低导致 DNA 气泡结构中 8-羟基鸟嘌呤的切口减少。总之，我们的数据支持 CSB 和 NEIL2 在转录相关碱基切除修复中的生物学作用。最后，使用全细胞提取物，CSB 敲低导致 DNA 气泡结构中 8-羟基鸟嘌呤的切口减少。总之，我们的数据支持 CSB 和 NEIL2 在转录相关碱基切除修复中的生物学作用。