Mutations in the Yellow Fever Virus Nonstructural Protein NS2A Selectively Block Production of Infectious Particles

Abstract

The study aims to demonstrate a possible link between bacterial cell metabolism and virulence by combining the metabolic mechanisms of the gram-negative bacteria, Escherichia coli. Glucose increases the proliferation of intestinal microflora, which augments the output of the short-chain fatty acids. Bacteria ferment glucose into short-chain fatty acids, which help regulate many biochemical processes and pathways. Each short-chain fatty acid maintains colonic pH, promotes cell differentiation, and the apoptosis of colonocytes. The long-chain fatty acids are also synthesized for plasma membrane and biofilm formation. To increase the synthesis of acetyl-CoA carboxylase, an enzyme that catabolizes glucose into short- and long-chain fatty acids, Escherichia coli was cultured in Luria broth enhanced with a high to a low concentration of glucose. The 15mM, a high concentration of glucose, yielded qPCR products measured for the target gene accA, which was 4,210ng/µL. The 7.5mM sample produced a concentration equal to 375 ng/µL, and the control sample measured an accA concentration of 196 ng/µL. The gene accA, one of four subunits for the acetyl-CoA carboxylase enzyme, was suppressed by asRNA, producing a qPCR product of 63 gene copies. Antisense RNA for accA reduced the amount of Lux-S, a vital gene needed for propagating quorum-sensing signal molecules. The Lux-S gene, which is responsible for releasing autoinducer 2 for cell-to-cell quorum sensing, was reduced by the gene inhibition of accA with asRNA. The increase in Lux-S transcription augments biofilm production in support of spreading virulence. The implications of the study advocate for designing antibiotics that target bacterial cell metabolic processes to block bacterial antibiotic resistance.