Frontiers in Cell and Developmental Biology
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2025
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IF 4.3
Cheng Han,Shanshan Fu,Dachao Tang,Yuting Chen,Dan Liu,Zihao Feng,Yujie Gou,Chi Zhang,Weizhi Zhang,Leming Xiao,Jiayi Zhang,Cong Yi,Yu Xue,Di Peng
In Saccharomyces cerevisiae , Atg1 is a core autophagy-related (Atg) protein kinase (PK) in regulating macroautophagy/autophagy, by physically interacting with numerous other proteins, or by phosphorylating various substrates. It is unclear how many Atg1-interacting partners and substrates are also involved in regulating autophagy. Here, we conducted transcriptomic, proteomic and phosphoproteomic profiling of Atg1-dependent molecular landscapes during nitrogen starvation-triggered autophagy, and detected 244, 245 and 217 genes to be affected by ATG1 in the autophagic process at mRNA, protein, and phosphorylation levels, respectively. Based on the Atg1 interactome, we developed a novel artificial intelligence (AI) framework, inference of autophagy regulators from multi-omic data (iAMD), and predicted 12 Atg1-interacting partners and 17 substrates to be potentially functional in autophagy. Further experiments validated that Rgd1 and Whi5 are required for bulk autophagy, as well as physical interactions and co-localizations with Atg1 during autophagy. In particular, we demonstrated that 2 phosphorylation sites (p-sites), pS78 and pS149 of Whi5, are phosphorylated by Atg1 to regulate the formation of Atg1 puncta during autophagy initiation. A working model was illustrated to emphasize the importance of the Atg1-centered network in yeast autophagy. In addition, iAMD was extended to accurately predict Atg proteins and autophagy regulators from other PK interactomes, indicating a high transferability of the method. Taken together, we not only revealed new autophagy regulators from the Atg1 interactome, but also provided a useful resource for further analysis of yeast autophagy.
研究领域:
分子生物学
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生物信息学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL DENTAL JOURNAL
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2025
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IF 3.7
Jiadi Shen,Chenyu Weng,Shuxian Zhu,Wanjing Chen,Xuening Xiong,Xin Wei
Objectives Candida albicans ( C. albicans ) biofilms are well-known to be resistantto various antifungal agents. Autophagy is crucial for adapting to changes in nutrition conditions. Protein phosphatase 2A (PP2A) is prominent in regulating physiological processes, possibly related to autophagy-related (ATG) protein phosphorylation. This study hypothesizes that PP2A affects biofilm formation and drug resistance of C. albicans via autophagy induction. Materials and methods The expressions of PP2A catalytic subunit coding gene PPH21 were compared. The mutant strain ( pph21 Δ/Δ) was constructed and the biofilm was treated with autophagy activator (rapamycin, Rap). The biofilm formation, drug susceptibility and oxidative stress levels were examined. The autophagic activity was detected, along with the autophagosomes observed. The therapeutic efficacy of the antifungal agents was estimated on the mice model of C. albicans oral infection. Results PPH21 was associated with C. albicans biofilm formation and drug resistance. Autophagy activation by rapamycin can induce increased autophagy levels, while it was hindered in pph21 Δ/Δ. Besides, the protein levels of Atg13 and Atg1 were significantly down-regulated in pph21 Δ/Δ+Rap ( P < .01), along with its decreased regulatory capacity to oxidative stress. Autophagy activation can promote biofilm formation and improve drug resistance, while the absence of PPH21 may prevent the enhancement of drug resistance. Autophagy activation reduced the efficacy of antifungal agents in treating oral C. albicans infection in mice, among which pph21 Δ/Δ presented better therapeutic effects. Conclusion PP2A is important in the autophagy induction of C. albicans by participating in Atg13 phosphorylation, followed by Atg1 activation, further affecting its biofilm formation and drug resistance. Clinical relevance PP2A-induced autophagy may be a potential regulatory mechanism of C. albicans drug resistance. This appears to be a promising therapeutic strategy for managing C. albicans -related infectious diseases.
研究领域:
微生物学
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分子生物学
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药理学
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细胞生物学
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医学真菌学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOTECHNOLOGY
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2024
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IF 4.1
Gang Luo,Wen Yang,Zijian Geng,Yiyi Cheng,Yingqing Xu,Yimeng Xiao,Jiying Liu
The Pichia pastoris expression system is a favorable platform for production of pharmaceutical proteins. Treatment of strains with N-acetyl-L-cysteine (NAC) has been shown to enhance the yield of recombinant proteins, thereby contributing to a reduction in production costs. However, the specific mechanism of action of NAC remains unclear. Previous research has indicated that glutathione (GSH) and autophagy are involved in the increased production of human serum albumin and porcine follicle-stimulating hormone β (HSA-pFSHβ) by NAC. This study investigated the potential interaction between GSH and autophagy in the production of HSA-pFSHβ. The findings indicated that sulfhydryl-free antioxidants such as melatonin, vitamin C, or vitamin E did not exhibit similar effects to NAC in enhancing HSA-pFSHβ yield. Moreover, NAC was found to enhance HSA-pFSHβ production by modulating GSH metabolism to reduce GSH consumption, increase total GSH levels, as well as glutathione peroxidase (GSH-Px) and glutathione reductase (GR) activities. Additionally, inhibition of autophagy through disruption of autophagy scaffolding proteins Atg1 or Atg11 led to an increase in recombinant HSA-pFSHβ production. Furthermore, NAC significantly decreased the phosphorylation of Slt2, and the absence of the SLT2 gene influenced the effect of NAC on HSA-pFSHβ secretion by modulating mitophagy and GSH metabolism. In conclusion, these results suggest a complex interplay between GSH metabolism and autophagy in the regulation of NAC-induced HSA-pFSHβ secretion.
研究领域:
分子生物学
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生物技术
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药用蛋白质
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酵母表达系统
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）