Aspergillus fumigatus Elongator complex subunit 3 affects hyphal growth, adhesion and virulence through wobble uridine tRNA modification.
Aspergillus fumigatus Elongator complex subunit 3 affects hyphal growth, adhesion and virulence through wobble uridine tRNA modification.
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烟曲霉 Elongator 复合物亚基 3 通过摆动尿苷 tRNA 修饰影响菌丝生长、粘附和毒力
DOI：
10.1371/journal.ppat.1010976
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发表时间：
2022-11
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
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作者：
真核多亚基Elongator复合物在转录延伸、组蛋白乙酰化和tRNA修饰中具有多种功能。然而，延长复合体在不同的生物体中起着不同的作用，其潜在的机制尚未被探索。此外，在人类真菌病原体中的延伸复合物的生物学功能仍然未知。在这项研究中，我们验证了条件致病真菌烟曲霉的延伸复合体由6个亚基（Elp 1 -6）组成，任何亚基的损失导致类似的缺陷菌落表型与受损的菌丝生长和减少分生孢子。Elongator复合物的催化亚基Elp 3包括一个S-腺苷甲硫氨酸结合（rSAM）结构域和一个赖氨酸乙酰转移酶（KAT）结构域，它在A.然而，Elp 3不影响体内H3 K14乙酰化水平。LC-MS/MS分析表明，Elp 3的缺失消除了tRNA摆动尿苷（U34）的5-甲氧羰基甲基-2-硫代尿苷（mcm 5s 2U）修饰，而通常含有mcm 5s 2U修饰的tRNAGlnUUG和tRNAGluUUC的过表达主要挽救了Δ elp 3突变体的缺陷，表明在A.烟熏。引人注目的是，全球蛋白质组学比较分析显示，与野生型菌株相比，Δ elp 3突变菌株中与氨基酸代谢相关的基因表达显著上调。Western blotting结果显示，缺失elp 3导致了氨基酸饥饿反应转录因子CpcA的过度表达，CpcA的缺失显著逆转了Δ elp 3突变体的缺陷表型，包括减弱了毒力。因此，本研究结果表明，A.烟曲霉Elp 3作为一种tRNA修饰酶，通过维持细胞内氨基酸稳态来调节生长、GAG产生、粘附和毒力。更广泛地说，我们的研究强调了U34 tRNA修饰在调节真菌病原体的细胞代谢状态和毒力性状中的重要性。
The eukaryotic multisubunit Elongator complex has been shown to perform multiple functions in transcriptional elongation, histone acetylation and tRNA modification. However, the Elongator complex plays different roles in different organisms, and the underlying mechanisms remain unexplored. Moreover, the biological functions of the Elongator complex in human fungal pathogens remain unknown. In this study, we verified that the Elongator complex of the opportunistic fungal pathogen Aspergillus fumigatus consists of six subunits (Elp1-6), and the loss of any subunit results in similarly defective colony phenotypes with impaired hyphal growth and reduced conidiation. The catalytic subunit-Elp3 of the Elongator complex includes a S-adenosyl methionine binding (rSAM) domain and a lysine acetyltransferase (KAT) domain, and it plays key roles in the hyphal growth, biofilm-associated exopolysaccharide galactosaminogalactan (GAG) production, adhesion and virulence of A. fumigatus; however, Elp3 does not affect H3K14 acetylation levels in vivo. LC–MS/MS chromatograms revealed that loss of Elp3 abolished the 5-methoxycarbonylmethyl-2-thiouridine (mcm5s2U) modification of tRNA wobble uridine (U34), and the overexpression of tRNAGlnUUG and tRNAGluUUC, which normally harbor mcm5s2U modifications, mainly rescues the defects of the Δelp3 mutant, suggesting that tRNA modification rather than lysine acetyltransferase is responsible for the primary function of Elp3 in A. fumigatus. Strikingly, global proteomic comparison analyses showed significantly upregulated expression of genes related to amino acid metabolism in the Δelp3 mutant strain compared to the wild-type strain. Western blotting showed that deletion of elp3 resulted in overexpression of the amino acid starvation-responsive transcription factor CpcA, and deletion of CpcA markedly reversed the defective phenotypes of the Δelp3 mutant, including attenuated virulence. Therefore, the findings of this study demonstrate that A. fumigatus Elp3 functions as a tRNA-modifying enzyme in the regulation of growth, GAG production, adhesion and virulence by maintaining intracellular amino acid homeostasis. More broadly, our study highlights the importance of U34 tRNA modification in regulating cellular metabolic states and virulence traits of fungal pathogens.
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影响因子：
6.7
作者：
Gravelat FN;Beauvais A;Liu H;Lee MJ;Snarr BD;Chen D;Xu W;Kravtsov I;Hoareau CM;Vanier G;Urb M;Campoli P;Al Abdallah Q;Lehoux M;Chabot JC;Ouimet MC;Baptista SD;Fritz JH;Nierman WC;Latgé JP;Mitchell AP;Filler SG;Fontaine T;Sheppard DC
通讯作者：
Sheppard DC
影响因子：
3.6
作者：
Krappmann, S;Bignell, EM;Braus, GH
通讯作者：
Braus, GH
影响因子：
5.6
作者：
Jarosz M;Van Lijsebettens M;Woloszynska M
通讯作者：
Woloszynska M
影响因子：
4.8
作者：
Kojic M;Wainwright B
通讯作者：
Wainwright B
DOI：
10.3390/jof3040057
发表时间：
2017-10-18
期刊：
Journal of fungi (Basel, Switzerland)
影响因子：
--
作者：
Bongomin F;Gago S;Oladele RO;Denning DW
通讯作者：
Denning DW