Genetic contribution to high temperature tolerance in Cryptococcus neoformans.
Genetic contribution to high temperature tolerance in Cryptococcus neoformans.
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遗传对新型隐球菌耐高温性的贡献。
DOI：
10.1093/genetics/iyaa009
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发表时间：
2021
期刊：
影响因子：
3.3
通讯作者：
Kozubowski,Lukasz
中科院分区：
文献类型：
--
作者：
Stempinski,PiotrR;Zielinski,JessicaM;Dbouk,NadirH;Huey,ElizabethS;McCormack,EllenC;Rubin,AlexanderM;Chandrasekaran,Srikripa;Kozubowski,Lukasz
新型隐球菌（Cryptococcus neoformans）是一种重要的致病真菌，它依赖于一个复杂的信号网络来适应和生存于宿主温度。蛋白磷酸酶钙调磷酸酶在37°C下对增殖至关重要，但其确切贡献仍不明确。为了更好地定义基因对C.为了获得新形式的温度耐受性，在37°C和保持钙调磷酸酶失活的条件下筛选了4031个基因敲除的基因。随后对83种可能对37°C敏感的候选菌株进行了技术简单但稳健的测定，其中细胞暴露于温度梯度。这导致了46个基因的鉴定有助于最高温度，在此温度下C。新生儿可增殖（Tmax）。在抑制钙调磷酸酶的条件下，进一步评估了46种突变体的Tmax，这些突变体的特征在于一系列Tmaxon无药物培养基，这导致鉴定了几种先前未表征的敲除，这些敲除表现出与钙调磷酸酶抑制的合成相互作用。突变体，缺乏septin Cdc 11是那些最低的Tmax和未能增殖的钙调磷酸酶活性的情况下。为了进一步确定与钙调神经磷酸酶的联系以及septins在高温生长中的作用，测试了46种突变体在37°C下的细胞形态以及在存在破坏细胞壁和细胞膜的试剂的情况下的生长。对钙调神经磷酸酶抑制敏感的突变体进行了测试，用于缺失编码septin的CDC 12和定位septin Cdc 3-mCherry的合成致死相互作用。这里描述的分析指出，以前未表征的基因，在标准的生长试验中错过了温度梯度试验是一个有价值的补充工具，用于阐明微生物增殖的温度范围的遗传基础。
The human fungal pathogenCryptococcus neoformansrelies on a complex signaling network for the adaptation and survival at the host temperature. Protein phosphatase calcineurin is central to proliferation at 37°C but its exact contributions remain ill-defined. To better define genetic contributions to theC. neoformanstemperature tolerance, 4031 gene knockouts were screened for genes essential at 37°C and under conditions that keep calcineurin inactive. Identified 83 candidate strains, potentially sensitive to 37°C, were subsequently subject to technologically simple yet robust assay, in which cells are exposed to a temperature gradient. This has resulted in identification of 46 genes contributing to the maximum temperature at whichC. neoformanscan proliferate (Tmax). The 46 mutants, characterized by a range of Tmaxon drug-free media, were further assessed for Tmaxunder conditions that inhibit calcineurin, which led to identification of several previously uncharacterized knockouts exhibiting synthetic interaction with the inhibition of calcineurin. A mutant that lacked septin Cdc11 was among those with the lowest Tmaxand failed to proliferate in the absence of calcineurin activity. To further define connections with calcineurin and the role for septins in high temperature growth, the 46 mutants were tested for cell morphology at 37°C and growth in the presence of agents disrupting cell wall and cell membrane. Mutants sensitive to calcineurin inhibition were tested for synthetic lethal interaction with deletion of the septin-encodingCDC12and the localization of the septin Cdc3-mCherry. The analysis described here pointed to previously uncharacterized genes that were missed in standard growth assays indicating that the temperature gradient assay is a valuable complementary tool for elucidating the genetic basis of temperature range at which microorganisms proliferate.
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影响因子：
1.5
作者：
Michael S. Waugh;Michael S. Waugh;Connie B Nichols;Connie B Nichols;Cameron M DeCesare;G. Cox;J. Heitman;J. Alspaugh
通讯作者：
Michael S. Waugh;Michael S. Waugh;Connie B Nichols;Connie B Nichols;Cameron M DeCesare;G. Cox;J. Heitman;J. Alspaugh
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影响因子：
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作者：
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影响因子：
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作者：
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DOI：
10.1021/acs.langmuir.6b03452
发表时间：
2016
期刊：
Langmuir : the ACS journal of surfaces and colloids
影响因子：
--
作者：
Shunsuke Yamada;Takumi Isogai;R. Tero;Y. Tanaka;T. Ujihara;M. Kinoshita;K. Takiguchi
通讯作者：
K. Takiguchi
影响因子：
3.6
作者：
Fox, DS;Cruz, MC;Heitman, J
通讯作者：
Heitman, J