PMID: 11918677Mar 29, 2002Paper

Detection of tmRNA-mediated trans-translation products in Bacillus subtilis

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
Ai FujiharaAkira Muto

Abstract

Bacterial tmRNA (10Sa RNA) is involved in a trans-translation reaction, which contributes to the degradation of incompletely synthesized peptides and the recycling of stalled ribosomes. To investigate the physiological roles of this reaction in Bacillus subtilis, we devised a system for detecting the proteins that are subject to in vivo trans-translation. The wild-type tmRNA gene (ssrA) in the genome was replaced by a variant ssrA encoding a tag-peptide sequence containing six histidine residues (His-tag) and two aspartic acids at the C-terminus. The His-tagged proteins that accumulated in the cells without degradation were fractionated by Ni2+-NTA column and gel electrophoresis and were detected by Western blotting with an anti-His-tag antibody. The results showed that the trans-translation occurred more frequently at a high temperature (50 degrees C) than at a low temperature (37 degrees C). Two-dimensional (2D) gel electrophoresis of the products revealed many distinct spots, which represent specific target proteins for the trans-translation reaction. Furthermore, the 2D gel patterns of the products from cells cultured at high and low temperatures were apparently different. Several tagged proteins were identified by the N-te...Continue Reading

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Citations

Aug 6, 2004·Nucleic Acids Research·Takayuki KonnoHyouta Himeno
Oct 6, 2005·Nucleic Acids Research·Krisana AsanoHyouta Himeno
Oct 26, 2007·Nucleic Acids Research·Daisuke KuritaHyouta Himeno
Apr 5, 2005·Journal of Bacteriology·Irina A OussenkoDavid H Bechhofer
Nov 2, 2006·RNA·Preeti MehtaA Wali Karzai
Dec 15, 2010·PloS One·Zachary C Ruhe, Christopher S Hayes
Oct 18, 2007·Proceedings of the National Academy of Sciences of the United States of America·Sue-Jean HongKenneth C Keiler
Nov 17, 2009·FEBS Letters·Christopher S Hayes, Kenneth C Keiler
Mar 27, 2007·Current Opinion in Microbiology·Kenneth C Keiler
Aug 5, 2008·Molecular Microbiology·Teppei AbeAkira Muto
Oct 1, 2011·Wiley Interdisciplinary Reviews. RNA·Sharief BarendsGilles P van Wezel
Dec 3, 2014·Biochimie·Hyouta HimenoTatsuhiko Abo
May 6, 2003·Annual Review of Microbiology·Jeffrey H Withey, David I Friedman
Apr 30, 2014·Frontiers in Genetics·Hyouta HimenoAkira Muto
Feb 10, 2004·Microbiology·Marc A B Kolkman, Eugenio Ferrari
May 20, 2003·The Journal of Biological Chemistry·Toshiharu TakahashiHyouta Himeno
Aug 28, 2020·Frontiers in Molecular Biosciences·Inam Ul HaqSabine Brantl

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