Lysine Succinylation Contributes to Aflatoxin Production and Pathogenicity in Aspergillus flavus

Abstract

Aspergillus flavus (A. flavus) is a ubiquitous saprophytic and pathogenic fungus that produces the aflatoxin carcinogen, and A. flavus can have tremendous economic and health impacts worldwide. Increasing evidence demonstrates that lysine succinylation plays an important regulatory role in metabolic processes in both bacterial and human cells. However, little is known about the extent and function of lysine succinylation in A. flavus. Here, we performed a global succinylome analysis of A. flavus using high accuracy nano-LC-MS/MS in combination with the enrichment of succinylated peptides from digested cell lysates and subsequent peptide identification. In total, 985 succinylation sites on 349 succinylated proteins were identified in this pathogen. Bioinformatics analysis revealed that the succinylated proteins were involved in various biological processes and were particularly enriched in the aflatoxin biosynthesis process. Site-specific mutagenesis and biochemical studies showed that lysine succinylation on the norsolorinic acid reductase NorA (AflE), a key enzyme in aflatoxins biosynthesis, can affect the production of sclerotia and aflatoxins biosynthesis in A. flavus. Together, our findings reveal widespread roles for lysine succinylation in regulating metabolism and aflatoxins biosynthesis in A. flavus. Our data provide a rich resource for functional analyses of lysine succinylation and facilitate the dissection of metabolic networks in this pathogen.

Lysine acetylation contributes to development, aflatoxin biosynthesis and pathogenicity in Aspergillus flavus

“…In A. flavus , Lan found that histone acetyltransferase and deacetylase play important roles in the production of aflatoxin by regulating modifications of histone acetylation (Lan et al ., ; Lan et al ., ). In addition, the results of acetylation‐mediated aflatoxin biosynthesis agree with the results of other reported PTMs in A. flavus , including phosphorylation (Ren et al ., ), SUMOylation (Nie et al ., ) and succinylation (Ren et al ., ). Taken together, we conclude that lysine acetylation is critical for secondary metabolism in A. flavus .…”

“…Previous studies have shown that histone deacetylase‐mediated acetylation modification regulates development, aflatoxin biosynthesis and pathogenicity of A. flavus (Lan et al ., ). In addition, proteomics analysis of phosphorylation and succinylation indicated that phosphorylation and succinylation regulate the development, pathogenicity and aflatoxin biosynthesis of A. flavus by regulating the function of key proteins (Ren et al ., ; ). Therefore, we believe that acetylation plays an important role in regulating various processes, and the model is suitable for the results presented in our study.…”

Integrated Succinylome and Metabolome Profiling Reveals Crucial Role of S-Ribosylhomocysteine Lyase in Quorum Sensing and Metabolism of Aeromonas hydrophila*

“…Enrichment of Lysine-succinylated Peptides-To enrich for a high abundance of lysine-succinylated peptides, immunoaffinity analysis was performed using agarose-conjugated anti-succinyllysine antibody (PTM Biolabs Inc., Hangzhou, China), as previously described (27). Briefly, the tryptic digested peptides were incubated with antisuccinyllysine agarose beads overnight at 4°C in NETN buffer (100 mM NaCl, 50 mM Tris-HCl, 1 mM EDTA, and 0.5% Nonidet P-40, pH 8.0), and then eluted with 1% trifluoroacetic acid (TFA).…”

Large-Scale Assessment of Bioinformatics Tools for Lysine Succinylation Sites

“…It should, however, be noted that the succinylation proteins overlap with other modifications due to some exhibiting dual properties. Recently many studies have suggested that lysine succinylation extensively overlaps with acetylation [25,27,42,63,68,73,74,75,76].…”

“…This modification can make notable alterations in protein function and structure regulation [3,13,35,36,37]. It can also participate in regulating many biological processes such as calorie restriction and metabolisms [38,39,40,41,42,43,44]. The identification of protein succinylation sites is a crucial topic in cellular pathology and physiology, which may provide valuable information for biomedical research and drug development.…”