PMID:7533896

Citation	McDowall, KJ, Kaberdin, VR, Wu, SW, Cohen, SN and Lin-Chao, S (1995) Site-specific RNase E cleavage of oligonucleotides and inhibition by stem-loops. Nature 374:287-90
Abstract	The enzyme RNase E (ref. 1) cuts RNA at specific sites within single-stranded segments. The role of adjacent regions of secondary structure in such cleavages is controversial. Here we report that 10-13-nucleotide oligomers lacking any stem-loop but containing the RNase E-cleaved sequence of RNA I, the antisense repressor of replication of ColE1-type plasmids, are cut at the same phosphodiester bond as, and 20 times more efficiently than, RNA I. These findings indicate that, contrary to previous proposals, stem-loops do not serve as entry sites for RNase E, but instead limit cleavage at potentially susceptible sites. Cleavage was reduced further by mutations in a non-adjacent stem-loop, suggesting that distant conformational changes can also affect enzyme access. Modulation of RNase E cleavages by stem-loop regions and to a lesser extent by higher-order structure may explain why this enzyme, which does not have stringent sequence specificity, cleaves complex RNAs at a limited number of sites.
Links	PubMed Online version:10.1038/374287a0
Keywords	Base Sequence; Endoribonucleases/antagonists & inhibitors; Endoribonucleases/metabolism; Escherichia coli/enzymology; Molecular Sequence Data; Mutation; Nucleic Acid Conformation; Oligoribonucleotides/metabolism; RNA, Bacterial/chemistry; RNA, Bacterial/genetics; RNA, Bacterial/metabolism; Ribonuclease T1/metabolism; Substrate Specificity
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