Han Liantao, Wu Yujuan, Xiong Sichu, Liu Tongbao
State Key Laboratory of Resource Insects, Southwest University Medical Research Institute, Chongqing 400715, China.
Jinfeng Laboratory, Chongqing 401329, China.
J Fungi (Basel). 2023 Jun 24;9(7):699. doi: 10.3390/jof9070699.
F-box protein is a key protein of the SCF E3 ubiquitin ligase complex, responsible for substrate recognition and degradation through specific interactions. Previous studies have shown that F-box proteins play crucial roles in sexual reproduction. However, the molecular mechanism by which F-box proteins regulate sexual reproduction in is unclear. In the study, we discovered the AICAR transformylase/IMP cyclohydrolase Ade16 as a substrate of Fbp1. Through protein interaction and stability experiments, we demonstrated that Ade16 is a substrate for Fbp1. To examine the role of in , we constructed the i strains and strains to analyze the function of Ade16. Our results revealed that the i strains had a smaller capsule and showed growth defects under NaCl, while the strains were sensitive to SDS but not to Congo red, which is consistent with the stress phenotype of the Δ strains, indicating that the intracellular protein expression level after overexpression was similar to that after deletion. Interestingly, although i strains can produce basidiospores normally, strains can produce mating mycelia but not basidiospores after mating, which is consistent with the Δmutant strains, suggesting that Fbp1 is likely to regulate the sexual reproduction of through the modulation of Ade16. A fungal nuclei development assay showed that the nuclei of the strains failed to fuse in the bilateral mating, indicating that Ade16 plays a crucial role in the regulation of meiosis during mating. In summary, our findings have revealed a new determinant factor involved in fungal development related to the post-translational regulation of AICAR transformylase/IMP cyclohydrolase.
F-box蛋白是SCF E3泛素连接酶复合体的关键蛋白，负责通过特异性相互作用识别和降解底物。先前的研究表明，F-box蛋白在有性生殖中发挥着关键作用。然而，F-box蛋白调节有性生殖的分子机制尚不清楚。在本研究中，我们发现AICAR转甲酰基酶/IMP环水解酶Ade16是Fbp1的底物。通过蛋白质相互作用和稳定性实验，我们证明Ade16是Fbp1的底物。为了研究Ade16在中的作用，我们构建了i菌株和菌株来分析Ade16的功能。我们的结果显示，i菌株的荚膜较小，在NaCl条件下表现出生长缺陷，而菌株对SDS敏感，但对刚果红不敏感，这与Δ菌株的应激表型一致，表明过表达后的细胞内蛋白质表达水平与缺失后相似。有趣的是，尽管i菌株能正常产生担孢子，但菌株在交配后能产生交配菌丝体但不能产生担孢子，这与Δ突变菌株一致，表明Fbp1可能通过调节Ade16来调节的有性生殖。一项真菌细胞核发育分析表明，菌株的细胞核在双边交配中未能融合，表明Ade16在交配过程中减数分裂的调节中起关键作用。总之，我们的研究结果揭示了一个与AICAR转甲酰基酶/IMP环水解酶的翻译后调控相关的参与真菌发育的新决定因素。
