PMID: 8609612Feb 16, 1996Paper

Two competing pathways for self-splicing by group II introns: a quantitative analysis of in vitro reaction rates and products

Journal of Molecular Biology
D L DanielsA M Pyle

Abstract

Self-splicing group II introns are found in bacteria and in the organellar genes in plants, fungi, and yeast. The mechanism for the first step of splicing is generally believed to involve attack of a specific intronic 2'-hydroxyl group on a phosphodiester linkage at the 5'-splice site, resulting in the formation of a lariat intron species. In this paper, we present kinetic and enzymatic evidence that in vitro there are two distinct pathways for group II intron self-splicing: one involves 2'-OH attack and another involves attack of water or hydroxide. These two pathways occur in parallel under all reaction conditions, although either can dominate in the presence of particular salts or protein cofactors. Both pathways are followed by a successful second step of splicing, and either pathway can be highly efficient. We find that the hydrolytic pathway prevails under physiological ionic conditions, while branching predominates at molar concentrations of ammonium ion. The intron is observed to adopt two major active conformations. In order to quantify their individual reaction rates, we applied a mechanistic model describing biphasic parallel kinetic behavior. Kinetic analysis throughout the investigation reveals that there is no cou...Continue Reading

Citations

Mar 1, 2006·Proceedings of the National Academy of Sciences of the United States of America·Sabine MohrAlan M Lambowitz
Nov 9, 2011·Molecular Genetics and Genomics : MGG·Zhi Cheng, Thomas M Menees
Jul 9, 2016·Annual Review of Biophysics·Anna Marie Pyle
Sep 5, 2008·PloS One·Marta MastroianniAlan M Lambowitz
Feb 2, 2010·Nature Chemical Biology·Michael RoitzschAnna Marie Pyle
Jun 18, 2010·Nucleic Acids Research·Nora ZinglerAnna Marie Pyle
Jan 4, 2013·PloS One·Jean-Claude FarréBenoît Castandet
Oct 17, 2018·Nature Chemical Biology·Olga FedorovaAnna Marie Pyle
Dec 25, 2019·Proceedings of the National Academy of Sciences of the United States of America·Alfredo J HernandezJeannie T Lee
Mar 12, 1998·Nature·M PodarP S Perlman
Sep 18, 2004·Proceedings of the National Academy of Sciences of the United States of America·Natalia G StarostinaMichael P Terns
May 8, 2010·Critical Reviews in Biochemistry and Molecular Biology·Anna Marie Pyle
Nov 10, 2013·Nucleic Acids Research·Srinivas SomarowthuAnna Marie Pyle
Nov 19, 2015·Scientific Reports·Ulrike Pfreundt, Wolfgang R Hess
Jun 16, 2007·Biological Chemistry·Olga Fedorova, Nora Zingler
Mar 24, 2009·Nucleic Acids Research·Fredrik B StabellAnne-Brit Kolstø
May 20, 2011·Chemistry : a European Journal·Tuomas Lönnberg
Oct 7, 2020·Nucleic Acids Research·Nan LiuMarlene Belfort
Aug 24, 2000·Biochemistry·A Bar-Shalom, M J Moore
Apr 4, 1997·Journal of Molecular Biology·M CostaF Michel
Jun 26, 1998·Journal of Molecular Biology·A R Ferré-D'AmaréJ A Doudna
Jul 17, 1998·Current Opinion in Structural Biology·P Z Qin, A M Pyle
Nov 15, 2002·Methods : a Companion to Methods in Enzymology·Olga FedorovaAnna Marie Pyle
Jul 14, 1998·Molecular Cell·B B KonfortiA M Pyle
Jun 23, 2021·Nucleic Acids Research·Félix LaRoche-JohnstonBenoit Cousineau

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