Analysis of the structure and folding of the 3' genomic RNA of flaviviruses

The structure of the 3' genomic RNA of flaviviruses has been analyzed by thermal melting, and by chemical and enzymatic probing of model RNAs. The 3' RNA had more transitions than could be assigned to individual stem-loops and a transition in the 40-45 degrees C range was assigned to tertiary structure unfolding. In support of this assignment, mutants designed to destabilize the proposed pseudoknot tertiary structure lose the 40-45 degrees C transition. Enzyme cleavage of the RNA and chemical probing as a function of temperature also support a pseudoknot tertiary structure for the 3' flavivirus RNA.