The impact of lysine succinylation modification of host factor for RNA phage Qβ replicase at K56 site on the biological functions of Aeromonas hydrophila.

Post-translational modifications (PTMs) play pivotal roles in modulating protein functions. In Aeromonas hydrophila, a freshwater Gram-negative pathogen threatening human and animal health, the host factor for RNA phage Qβ replicase (Hfq) is a key regulatory protein. Its K56 site undergoes succinylation modification, suggesting important functions. However, the exact roles of Hfq and its K56 site in A. hydrophila were unclear. Our physiological assays showed that hfq deletion increased protease activity and motility, but reduced biofilm formation, chemotaxis, stress response, and antibiotic resistance. K56 succinylation enhanced protease activity and biofilm formation, and altered chemotaxis. Comprehensive transcriptome and proteome analysis further revealed that hfq deletion led to an increase in 11 peptidases and 8 flagellar proteins, as well as a decrease in 5 stress-related proteins. It also suppressed the pathways involved in chemotaxis and beta-lactam resistance. On the other hand, K56 succinylation enhanced fatty acid synthesis pathway and the expression of methyl-accepting chemotaxis proteins (MCPs). Consistent with the omics data, qPCR validation confirmed the expression of 12 genes, including metalloprotease and MCPs. Overall, this study deepened insights into Hfq regulation in A. hydrophila and revealed the previously unknown molecular switch function of post-translational modifications (PTMs) on this protein.