A single mutation in the gatekeeper residue in TgMAPKL-1 restores the inhibitory effect of a bumped kinase inhibitor on the cell cycle.
A single mutation in the gatekeeper residue in TgMAPKL-1 restores the inhibitory effect of a bumped kinase inhibitor on the cell cycle.
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DOI：
10.1016/j.ijpddr.2014.12.001
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发表时间：
2015-04
影响因子：
4
通讯作者：
Kato, Kentaro
中科院分区：
文献类型：
--
作者：
Sugi, Tatsuki;Kawazu, Shin-ichiro;Horimoto, Taisuke;Kato, Kentaro
TgMAPKL-1中Ser191的替换改变了弓形虫对突变的激酶抑制剂的敏感性。抑制TgMAPKL-1可导致宿主体内寄生虫细胞增大。放大的寄生虫复制DNA，但不能完成胞质分裂。抑制TgMAPKL-1抑制子代细胞的萌发。弓形虫是弓形虫病的病原体。蛋白激酶抑制剂1 nm-PP1通过靶向TgCDPK1抑制弓形虫的生长。然而，我们最近报道，对1 nm-PP1的抗性可以通过弓形虫有丝分裂原激活的蛋白激酶样1(TgMAPKL-1)的突变来获得。进一步鉴定TgMAPKL-1突变如何恢复1 nm-PP1的抑制作用，将进一步阐明TgMAPKL-1在寄生虫生活史中的功能。因此，我们用TgMAPKL-1突变在守门人残基Ser191上的寄生虫克隆，这是1 nm-PP1敏感性的关键。RH/ku80-/HA-TgMAPKL-1S191Y的宿主细胞裂解在250 nm处被完全抑制，而RH/ku80-/HA-TgMAPKL-1S191Y的裂解不受抑制。通过比较1 nm-PP1敏感克隆(RH/ku80-/HA-TgMAPKL-1S191A)和抗性克隆(RH/ku80-/HA-TgMAPKL-1S191Y)，我们观察到抑制TgMAPKL-1抑制DNA复制后细胞周期进展。形态学分析显示，抑制TgMAPKL-1可导致寄生虫细胞增大，子代细胞支架增多，胞质分裂不完全。结论：TgMAPKL-1基因突变恢复了1 nm-PP1对弓形虫内源性细胞周期的抑制作用。鉴于内生作用是该寄生虫速殖子和缓殖子阶段细胞分裂的主要机制，TgMAPKL-1可能是一个有希望的药物开发靶点。对调控TgMAPKL-1的信号的探索将为进一步深入了解弓形虫独特的细胞分裂模式提供依据。
Substitution of Ser191 in TgMAPKL-1 changed T. gondii susceptibility to bumped kinase inhibitors. Inhibition of TgMAPKL-1 caused enlarged parasite cells in the host. Enlarged parasites replicated DNA but failed to complete cytokinesis. Inhibition of TgMAPKL-1 arrested the budding of daughter cells. Toxoplasma gondii is the causative pathogen for Toxoplasmosis. Bumped kinase inhibitor 1NM-PP1 inhibits the growth of T. gondii by targeting TgCDPK1. However, we recently reported that resistance to 1NM-PP1 can be acquired via a mutation in T. gondii mitogen-activated protein kinase like 1 (TgMAPKL-1). Further characterization of how this TgMAPKL-1 mutation restores the inhibitory effect of 1NM-PP1 would shed further light on the function of TgMAPKL-1 in the parasite life cycle. Therefore, we made parasite clones with TgMAPKL-1 mutated at the gatekeeper residue Ser 191, which is critical for 1NM-PP1 susceptibility. Host cell lysis of RH/ku80-/HA-TgMAPKL-1S191A was completely inhibited at 250 nM 1NM-PP1, whereas that of RH/ku80-/HA-TgMAPKL-1S191Y was not. By comparing 1NM-PP1-sensitive (RH/ku80-/HA-TgMAPKL-1S191A) and -resistant (RH/ku80-/HA-TgMAPKL-1S191Y) clones, we observed that inhibition of TgMAPKL-1 blocked cell cycle progression after DNA duplication. Morphological analysis revealed that TgMAPKL-1 inhibition caused enlarged parasite cells with many daughter cell scaffolds and imcomplete cytokinesis. We conclude that the mutation in TgMAPKL-1 restored the cell cycle-arresting effect of 1NM-PP1 on T. gondii endodyogeny. Given that endodyogeny is the primary mechanism of cell division for both the tachyzoite and bradyzoite stages of this parasite, TgMAPKL-1 may be a promising target for drug development. Exploration of the signals that regulate TgMAPKL-1 will provide further insights into the unique mode of T. gondii cell division.
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影响因子：
3.4
作者：
Morlon-Guyot J;Berry L;Chen CT;Gubbels MJ;Lebrun M;Daher W
通讯作者：
Daher W
影响因子：
64.8
作者：
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影响因子：
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作者：
Gubbels MJ;Lehmann M;Muthalagi M;Jerome ME;Brooks CF;Szatanek T;Flynn J;Parrot B;Radke J;Striepen B;White MW
通讯作者：
White MW
影响因子：
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作者：
Miranda-Saavedra, Diego;Gabaldon, Toni;Doerig, Christian
通讯作者：
Doerig, Christian
影响因子：
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作者：
Sugi, Tatsuki;Kato, Kentaro;Akashi, Hiroomi
通讯作者：
Akashi, Hiroomi