SHMT, SHMTML and PRPS1 synergize to regulate blood digestion and nutrient metabolism in Aedes aegypti mosquitoes

Mosquitoes rely on vertebrate blood for nutrients vital for ovarian development. The enzyme serine hydroxymethyltransferase (SHMT) is crucial for amino acid and one‑carbon metabolism, playing a significant role in blood digestion and nutrient use in mosquitoes, though its functional mechanism remains further investigation. Using CRISPR/Cas9 to knock out the SHMT gene, we observed impaired blood digestion, delayed ovarian development, and inability to fly in mosquitoes. Multi-omics analysis revealed that SHMT deletion affected genes and metabolites related to amino acid metabolism. Knocking down SHMT-responsive genes mitochondrial-like serine hydroxymethyltransferase (SHMTML) and ribose-phosphate pyrophosphokinase 1 (PRPS1) also hindered blood digestion and ovarian development, mirroring SHMT-deficient mosquitoes. The interaction between SHMT, SHMTML, and PRPS1 was confirmed through various experiments, including Co-IP, GST pull-down, immunofluorescence colocalization, BiFC, molecular docking, and functional studies. Further research reveals that missing any of these proteins in mosquitoes results in ammonia and reactive oxygen species buildup, leading to mitochondrial problems, midgut cell damage, and abnormal enzyme expression. This study highlights a new molecular mechanism of SHMT and emphasizes its crucial interaction with SHMTML and PRPS1 in blood digestion and nutrient metabolism in vector mosquitoes. These findings may offer a strategic foundation for the development of innovative mosquito control measures.