Residues in Escherichia coli RNase P RNA important for cleavage site selection and divalent metal ion binding

Journal of Molecular Biology
J Kufel, L A Kirsebom

Abstract

We have used genetics as a tool to study the importance of an internal loop (P7) of Escherichia coli RNase P RNA (M1 RNA) in cleavage site selection and the binding of a divalent metal ion(s). The preferred cleavage site on a model tRNA precursor substrate shifted as a result of base-substitutions and deletions within this loop, in particular when changes were introduced at positions directly involved in base-pairing with the 3'-terminal RCCA motif of the substrate. Additionally, these changes in M1 RNA resulted in alterations in the binding of a divalent metal ion(s) in the vicinity of this internal loop as revealed by lead(II)-induced cleavage. From these data we conclude that the structural integrity of the P7 loop is important for both cleavage site selection and divalent metal ion binding. Cross-linking experiments using precursors carrying a 4-thioU immediately 5' of two independent cleavage sites suggest that close contact points between M1 RNA and nucleotides at these cleavage sites depend on the interaction between M1 RNA and the 3'-terminal RCCA motif of the substrate. Our findings further support the view that there are at least two different ways for the tRNA domain of a tRNA precursor to interact with M1 RNA. These...Continue Reading

Citations

Mar 10, 2001·Journal of Cellular Physiology·S XiaoD R Engelke
Dec 8, 2005·Nucleic Acids Research·Ema KikovskaLeif A Kirsebom
May 26, 1999·Proceedings of the National Academy of Sciences of the United States of America·N E MikkelsenL A Kirsebom
Oct 19, 2001·Proceedings of the National Academy of Sciences of the United States of America·M Brännvall, L A Kirsebom
Jun 9, 1999·Proceedings of the National Academy of Sciences of the United States of America·A Pascual, A Vioque
Sep 24, 1999·Journal of Molecular Biology·M Brännvall, L A Kirsebom
Apr 28, 2006·Journal of Molecular Biology·Ema KikovskaLeif A Kirsebom
Jul 12, 2005·Journal of Molecular Biology·Mathias Brännvall, Leif A Kirsebom
Apr 6, 2006·Critical Reviews in Biochemistry and Molecular Biology·Scott C Walker, David R Engelke
Jun 17, 1999·FEMS Microbiology Reviews·A Schön
Nov 30, 2002·Biochimie·Mathias BrännvallLeif A Kirsebom
Nov 15, 2002·Methods : a Companion to Methods in Enzymology·Eric L ChristianMichael E Harris
Aug 28, 2007·Journal of Molecular Biology·Mathias BrännvallLeif A Kirsebom

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