Position dependence of the rous sarcoma virus negative regulator of splicing element reflects proximity to a 5′ splice site

Abstract

Rous sarcoma virus (RSV) requires incomplete splicing of its viral transcripts to maintain efficient replication. A splicing inhibitor element, the negative regulator of splicing (NRS), is located near the 5′ end of the RNA but the significance of this positioning is not known. In a heterologous intron the NRS functions optimally when positioned close to the authentic 5′ splice site. This observation led us to investigate the basis of the position dependence. Four explanations were put forth and stressed the role of three major elements involved in splicing, the 3′ splice site, the 5′ splice site, and the 5′ end cap structure. NRS function was unrelated to its position relative to the 3′ splice site or the cap structure and appeared to depend on its position relative to the authentic 5′ splice site. We conclude that position dependence may reflect distance constraints necessary for competition of the NRS with the authentic 5′ splice site for pairing with the 3′ splice sites.