Genetic Interaction Analysis Reveals that Cryptococcus neoformans Utilizes Multiple Acetyl-CoA-Generating Pathways during Infection.
Genetic Interaction Analysis Reveals that Cryptococcus neoformans Utilizes Multiple Acetyl-CoA-Generating Pathways during Infection.
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DOI：
10.1128/mbio.01279-22
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发表时间：
2022-08-30
期刊：
影响因子：
6.4
通讯作者：
Krysan, Damian J.
中科院分区：
文献类型：
--
作者：
Alden, Katy M.;Jezewski, Andrew J.;Beattie, Sarah R.;Fox, David, III;Krysan, Damian J.
新型隐球菌是一种重要的人类真菌病原体，外部环境是其主要生态位。先前的工作表明，两种非必需的乙酰辅酶A代谢酶，ATP-柠檬酸裂解酶（ACL1）和乙酰辅酶A合成酶（ACS1），在新型隐球菌感染中发挥重要作用。在这里，我们采用遗传相互作用方法来研究这两种酶与最初称为 ACS2 的酶之间的相互作用，但我们发现它是乙酰乙酰辅酶 A 合成酶；我们根据其生化活性及其底物的系统名称，将基因重新命名为 2-酮丁酰 CoA 合成酶 1 (KBC1)。 ACL1 和 ACS1 代表基于我们的遗传相互作用研究的一对合成致死基因。 KBC1 与 ACS1 或 ACL1 的双突变体在体外不具有显着的合成表型，尽管我们发现删除这些酶中的任何一种都会降低巨噬细胞内的适应性。重要的是，相对于单突变体和 WT，acs1Δ kbc1Δ 双突变体在 CNS 中的适应性显着降低（真菌负荷减少约 2 log10 CFU），表明这些酶在感染过程中发挥的重要作用。与体外条件相比，体内 ACS1 和 KBC1 的表达均增加。 acs1Δ突变体在体内对氟康唑高度敏感，尽管其体外表型极小。这些数据不仅提供了对新型抗真菌 Acs 抑制剂的体内作用机制的深入了解，而且还提供了新型隐球菌对宿主环境的代谢适应。
Cryptococcus neoformans is an important human fungal pathogen for which the external environment is its primary niche. Previous work has shown that two nonessential acetyl-CoA metabolism enzymes, ATP-citrate lyase (ACL1) and acetyl-CoA synthetase (ACS1), play important roles in C. neoformans infection. Here, we took a genetic interaction approach to studying the interplay between these two enzymes along with an enzyme initially called ACS2 but which we have found is an acetoacetyl-CoA synthetase; we have renamed the gene 2-ketobutyryl CoA synthetase 1 (KBC1) based on its biochemical activity and the systematic name of its substrate. ACL1 and ACS1 represent a synthetic lethal pair of genes based on our genetic interaction studies. Double mutants of KBC1 with either ACS1 or ACL1 do not have significant synthetic phenotypes in vitro, although we find that deletion of any one of these enzymes reduces fitness within macrophages. Importantly, the acs1Δ kbc1Δ double mutant has significantly reduced fitness in the CNS relative to either single mutant as well as WT (~2 log10 CFU reduction in fungal burden), indicating the important role these enzymes play during infection. The expression of both ACS1 and KBC1 is increased in vivo relative to in vitro conditions. The acs1Δ mutant is hypersusceptible to fluconazole in vivo despite its minimal in vitro phenotypes. These data not only provide insights into the in vivo mechanism of action for a new class of antifungal Acs inhibitors but also into metabolic adaptations of C. neoformans to the host environment.
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