A 5′-3′ exonuclease activity involved in forming the 3′ products of histone pre-mRNA processing in vitro

Abstract

Histone RNA 3′ processing in vitro produces one or more 5′ cleavage products corresponding to the mature histone mRNA 3′ end, and a group of 3′ cleavage products whose 5′ ends are mostly located several nucleotides downstream of the mRNA 3′ end. The formation of these 3′ products is coupled to the formation of 5′ products and dependent on the U7 snRNP and a heat-labile processing factor. These short 3′ products therefore are a true and general feature of the processing reaction. Identical 3′ products are also formed from a model RNA containing all spacer nucleotides downstream of the mature mRNA 3′ end, but no sequences from the mature mRNA. Again, this reaction is dependent on both the U7 snRNP and a heat-labile factor. Unlike the processing with a full-length histone pre-mRNA, this reaction produces only 3′ but no 5′ fragments. In addition, product formation is inhibited by addition of cap structures at the model RNA 5′ end, indicating that product formation occurs by 5′-3′ exonucleolytic degradation. This degradation of a model 3′ product by a 5′-3′ exonuclease suggests a mechanism for the release of the U7 snRNP after processing by shortening the cut-off histone spacer sequences base paired to U7 RNA.