Characterization of the 3' terminal 42 nucleotide host protein binding element of the mouse hepatitis virus 3' untranslated region

Abstract

Mouse Hepatitis virus (MHV) is a member of the coronavirus family in the order Nidovirales. The 32 kb genome contains cis-acting sequences necessary for replication of the viral genome. Those cis-acting sequences have been shown to bind host proteins, and binding of those proteins is necessary for virus replication. One of the cis-acting elements is the 3' terminal 42 nucleotide host protein binding element. Previous work has demonstrated that mitochondrial aconitase, mitochondrial heat shock protein 70, heat shock protein 60 and heat shock protein 40 bind to the 3' terminal 42 nucleotide host protein binding element. We demonstrated that RNA secondary structure of the 3' terminal 42 nucleotide host protein binding element is necessary for host protein binding in vitro. We also demonstrate that primary structure of the 3' terminal 42 nucleotide host protein binding element is necessary for viral replication by targeted recombination. DI replication assays infer that the 3' terminal 42 nucleotide host protein binding element plays a role in positive strand synthesis from the negative strand template. Current studies involve the infectious cDNA clone, which will provide definitive answers on the role of the 3' terminal 42 nucleotide host protein binding element in MHV replication.