Essential roles for deubiquitination in Leishmania life cycle progression
Essential roles for deubiquitination in Leishmania life cycle progression
复制标题
去泛素化在利什曼原虫生命周期进展中的重要作用
DOI：
10.1101/2020.03.05.978528
复制
发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Damianou A
中科院分区：
文献类型：
--
作者：
Damianou A
寄生原生动物利什曼原虫需要蛋白酶体、自噬和溶酶体蛋白分解途径来执行其生命周期中发生的广泛的细胞重构。蛋白酶体对寄生虫的增殖是必不可少的，但对生长和分化过程中泛素化/去泛素化过程的要求却知之甚少。MicianaC12、C19和C65脱泛素化半胱氨酸肽酶(DUBS)的活性分析表明，在原环前鞭毛体向无鞭毛体分化的过程中，DUB活性保持相对恒定。然而，当对包括15个使用CRISPR-Cas9在前鞭毛体中创建的条形码Dub Null突变体的池进行生命周期表型(BAR-SEQ)时，在分化和细胞内感染期间观察到显著的适合性损失。DUB4、DUB7和DUB13是从超环前鞭毛体成功转化为无鞭毛体所必需的，DUB3、5、6、8、10、11和14是小鼠无鞭毛体正常增殖所必需的。DUB 1、DUB 2、DUB 12和DUB 16对前鞭毛体的存活是必不可少的，DUB 2在体内外的DiCre诱导基因缺失的实验中证明了DUB2在建立感染中的重要作用。DUB2位于细胞核中，与核蛋白相互作用，涉及转录/染色质动力学、mRNA剪接和mRNA封顶。DUB2具有广泛的连锁特异性，可切割除Lys27和MET1以外的所有二泛素链。我们的研究证明了DUBS在利什曼原虫的分化和细胞内生存中发挥的关键作用，并且无鞭毛体对泛素化动态平衡的破坏非常敏感。
The parasitic protozoanLeishmaniarequires proteasomal, autophagic and lysosomal proteolytic pathways to enact the extensive cellular remodelling that occurs during its life cycle. The proteasome is essential for parasite proliferation, yet little is known about the requirement for ubiquitination/deubiquitination processes in growth and differentiation. Activity-based protein profiling ofL.mexicanaC12, C19 and C65 deubiquitinating cysteine peptidases (DUBs) revealed DUB activity remains relatively constant during differentiation of procyclic promastigote to amastigote. However, when life cycle phenotyping (bar-seq) was performed on a pool including 15 barcoded DUB null mutants created in promastigotes using CRISPR-Cas9, significant loss of fitness was observed during differentiation and intracellular infection. DUBs 4, 7, and 13 are required for successful transformation from metacyclic promastigote to amastigote and DUBs 3, 5, 6, 8, 10, 11 and 14 are required for normal amastigote proliferation in mice. DUBs 1, 2, 12 and 16 are essential for promastigote viability and the essential role of DUB2 in establishing infection was demonstrated using DiCre inducible gene deletionin vitroandin vivo. DUB2 is found in the nucleus and interacts with nuclear proteins associated with transcription/chromatin dynamics, mRNA splicing and mRNA capping. DUB2 has broad linkage specificity, cleaving all the di-ubiquitin chains except for Lys27 and Met1. Our study demonstrates the crucial role that DUBs play in differentiation and intracellular survival ofLeishmaniaand that amastigotes are exquisitely sensitive to disruption of ubiquitination homeostasis.
登录
查看更多内容
影响因子：
6.7
作者：
Cayla M;Rachidi N;Leclercq O;Schmidt-Arras D;Rosenqvist H;Wiese M;Späth GF
通讯作者：
Späth GF
影响因子：
6.7
作者：
Saunders EC;Ng WW;Kloehn J;Chambers JM;Ng M;McConville MJ
通讯作者：
McConville MJ
影响因子：
4.9
作者：
Bringmann, Gerhard;Thomale, Katja;Schurigt, Uta
通讯作者：
Schurigt, Uta
利什曼原虫分化的调控动态：解卷积信号和识别磷酸化趋势*
影响因子：
7
作者：
P. Tsigankov;P. Gherardini;M. Helmer;G. Späth;P. Myler;D. Zilberstein
通讯作者：
D. Zilberstein
影响因子：
3.6
作者：
R. A. Williams;L. Tetley;J. Mottram;G. H. Coombs
通讯作者：
R. A. Williams;L. Tetley;J. Mottram;G. H. Coombs