Identification of three core regions essential for protein splicing of the yeast Vma1 protozyme. A random mutagenesis study of the entire Vma1-derived endonuclease sequence.

The Journal of Biological Chemistry
M KawasakiYasuhiro Anraku

Abstract

The translation product of the VMA1 gene of Saccharomyces cerevisiae undergoes protein splicing, in which the intervening region is autocatalytically excised and the franking regions are ligated. The splicing reaction is catalyzed essentially by the in-frame insert, VMA1-derived endonuclease (VDE), which is a site-specific endonuclease to mediate gene homing. Previous mutational analysis of the splicing reaction has been concentrated extensively upon the splice junctions. However, it still remains unknown which amino acid residues are crucial for the splicing reaction within the entire region of VDE and its neighboring elements. In this work, a polymerase chain reaction-based random mutagenesis strategy was used to identify such residues throughout the overall intervening sequence of the VMA1 gene. Splicing-defective mutant proteins were initially screened using a bacterial expression system and then analyzed further in yeast cells. Mutations were mapped at the N- and C-terminal splice junctions and around the N-terminal one-third of VDE. We identified four potent mutants that yielded aberrant products with molecular masses of 200, 90, and 80 kDa. We suggest that the conserved His362, newly identified as the essential residue f...Continue Reading

References

Dec 11, 1992·Nucleic Acids Research·R A HodgesW E Jack
Aug 1, 1992·PCR Methods and Applications·R C Cadwell, G F Joyce
Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·F B PerlerS K Hempstead
Dec 1, 1994·Protein Science : a Publication of the Protein Society·S Pietrokovski
Feb 1, 1994·Journal of Biochemistry·Y Anraku, R Hirata
Apr 16, 1996·Proceedings of the National Academy of Sciences of the United States of America·H FsihiS T Cole
May 24, 1996·Biochemical and Biophysical Research Communications·M KawasakiY Anraku

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Citations

Mar 26, 1998·Protein Science : a Publication of the Protein Society·S Pietrokovski
Sep 28, 1998·Biochimica Et Biophysica Acta·H WuX Q Liu
Aug 4, 2001·Trends in Genetics : TIG·S Pietrokovski
Jul 28, 2009·Journal of the American Chemical Society·Zhenming DuChunyu Wang
May 25, 2010·Nature Chemical Biology·Silvia FrutosTom W Muir
Oct 20, 2009·Proceedings of the National Academy of Sciences of the United States of America·Koji YaharaIchizo Kobayashi
Mar 28, 1998·The EMBO Journal·M W SouthworthF B Perler
Mar 26, 1999·Protein Science : a Publication of the Protein Society·S Daugelat, W R Jacobs
Feb 15, 2011·Journal of Bacteriology·Kazuo Tori, Francine B Perler
Oct 27, 2011·PloS One·Kazuo Tori, Francine B Perler
Nov 13, 2009·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Yasuhiro Anraku, Yoshinori Satow
Apr 17, 2004·Proceedings of the National Academy of Sciences of the United States of America·Alessandra RomanelliTom W Muir
Aug 5, 1998·Proceedings of the National Academy of Sciences of the United States of America·H WuX Q Liu
Aug 29, 2012·Cellular and Molecular Life Sciences : CMLS·Gerrit Volkmann, Henning D Mootz
Jun 5, 2007·FEBS Letters·Esther J PearlSigurd M Wilbanks
Jan 31, 2012·Fungal Genetics and Biology : FG & B·Annika A M BokorJan A L van Kan
Mar 17, 2006·IUBMB Life·Francine B Perler
Jun 4, 2010·Protein Science : a Publication of the Protein Society·Lear E BraceFrancine Perler
Nov 28, 2000·Annual Review of Genetics·X Q Liu
Aug 31, 2000·Annual Review of Biochemistry·H Paulus
Jun 1, 2011·Biomolecular Concepts·A Sesilja Aranko, Gerrit Volkmann
Jul 27, 2002·Annual Review of Microbiology·J Peter GogartenElena Hilario
Oct 6, 2016·Extremophiles : Life Under Extreme Conditions·Kazuo Tori, Francine Perler
Dec 12, 2017·Medical Mycology·Hans Garcia GarcesE Bagagli
Nov 20, 2018·Biotechnology and Bioengineering·C Seth PearsonGeorges Belfort
Apr 22, 2005·Molecular Biology and Evolution·Vassiliki Koufopanou, Austin Burt
Apr 4, 2014·The Journal of Biological Chemistry·Kenneth V MillsFrancine B Perler
Apr 20, 2000·The Journal of Biological Chemistry·L ChenF B Perler
Oct 11, 2015·The Journal of Biological Chemistry·Anne-Lena Bachmann, Henning D Mootz
Feb 7, 2004·European Journal of Biochemistry·Ralf DavidAnnette G Beck-Sickinger
Nov 8, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ervin KovácsZoltán Mucsi
Dec 20, 2020·Microorganisms·Ananya NandaSasmita Nayak
Jul 5, 2001·Bioorganic Chemistry·H Paulus
Jul 25, 2017·The Journal of Physical Chemistry. B·J I Mujika, X Lopez
Aug 3, 2013·Biochemistry·Michael C NicastriKenneth V Mills

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