Novel class of OTU deubiquitinases regulate substrate ubiquitination upon Legionella infection
Novel class of OTU deubiquitinases regulate substrate ubiquitination upon Legionella infection
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DOI：
10.1101/2020.04.25.060954
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发表时间：
2020-04
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通讯作者：
Donghyuk Shin;Anshu Bhattacharya;Yi-Lin Cheng;Marta Campos Alonso;A. Mehdipour;Gerbrand J. van der Heden van Noort-Gerbrand-J.-van-der-Heden-van-Noort-5581721;H. Ovaa;G. Hummer;I. Dikič
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作者：
Donghyuk Shin;Anshu Bhattacharya;Yi-Lin Cheng;Marta Campos Alonso;A. Mehdipour;Gerbrand J. van der Heden van Noort-Gerbrand-J.-van-der-Heden-van-Noort-5581721;H. Ovaa;G. Hummer;I. Dikič
嗜肺军团菌是一种导致军团病的革兰氏阴性致病菌。军团菌基因组编码 300 多种效应蛋白，能够在感染过程中调节宿主与病原体的相互作用。其中还有改变宿主泛素化系统的酶，包括细菌连接酶和去泛素酶。在本研究中，基于对 305 个军团菌效应蛋白的同源性检测筛选，我们鉴定了两种军团菌 OTU 样去泛素酶（LOT；LotB (Lpg1621/Ceg23) 和 LotC (Lpg2529)，LotA (Lpg2248/Lem21) 是已知的）。 LotC 催化核心 (LotC14-310) 的晶体结构在 2.4 Å 处确定，并与其他 OTU 去泛素酶（包括 LotB）进行比较。与经典 OTU 家族不同，军团菌 OTU 家族（LotB 和 LotC）的结构在 Cys 环和可变环之间显示出延伸的螺旋叶，这定义了一类新型 OTU 去泛素酶。尽管 LotB 和 LotC 的螺旋叶结构存在差异，但它们都与泛素相互作用。 LotB 具有一个额外的泛素结合位点 (S1’)，能够特异性切割 Lys63 连接的多聚泛素链。相比之下，LotC 仅包含 S1 位点并切割不同种类的泛素链。对催化失活 LotB 和 LotC 的 MS 分析确定了这两种相关 DUB 的不同类别的宿主底物。总之，我们的结果提供了细菌 OTU 去泛素酶的新结构见解，并表明细菌去泛素酶在宿主-病原体相互作用中的独特作用。
Legionella pneumophila is a gram-negative pathogenic bacterium that causes Legionaries’ disease. The Legionella genome codes more than 300 effector proteins able to modulate host-pathogen interactions during infection. Among them are also enzymes altering the host-ubiquitination system including bacterial ligases and deubiquitinases. In this study, based on homology-detection screening on 305 Legionella effector proteins, we identified two Legionella OTU-like deubiquitinases (LOT; LotB (Lpg1621/Ceg23) and LotC (Lpg2529), LotA (Lpg2248/Lem21) is already known). A crystal structure of LotC catalytic core (LotC14-310) was determined at 2.4 Å and compared with other OTU deubiquitinases, including LotB. Unlike the classical OTU-family, the structures of Legionella OTU-family (LotB and LotC) shows an extended helical lobe between the Cys-loop and the variable loop, which define a novel class of OTU-deubiquitinase. Despite structural differences in their helical lobes, both LotB and LotC interact with ubiquitin. LotB has an additional ubiquitin binding site (S1’) enabling specific cleavage of Lys63-linked poly-ubiquitin chains. By contrast, LotC only contains the S1 site and cleaves different species of ubiquitin chains. MS analysis of catalytically inactive LotB and LotC identified different categories of host-substrates for these two related DUBs. Together, our results provide new structural insights of bacterial OTU deubiquitinases and indicate distinct roles of bacterial deubiquitinases in host-pathogen interactions.