Zinc inhibition of protein trans-splicing and identification of regions essential for splicing and association of a split intein*

The Journal of Biological Chemistry
I GhoshM Q Xu

Abstract

Two important aspects of protein splicing were investigated by employing the trans-splicing intein from the dnaE gene of Synechocystis sp. PCC6803. First, we demonstrated that both protein splicing and cleavage at the N-terminal splice junction were inhibited in the presence of zinc ion. The trans-splicing reaction was partially blocked at a concentration of 1-10 microm Zn(2+) and completely inhibited at 100 microm Zn(2+); the inhibition by zinc was reversed in the presence of ethylenediaminetetraacetic acid. We propose that inactivation of Cys(160) at the C-terminal splice junction by the chelation of zinc affects both the N-S acyl rearrangement and the transesterification steps in the splicing pathway. Furthermore, in vivo and in vitro assays were established for the determination of intein residues and regions required for splicing or association between the N- and C-terminal intein halves. N-terminal truncation of the intein C-terminal segment inhibited both splicing and association activities, suggesting this region is crucial for the formation of an interface between the two intein halves. The replacement of conserved residues in blocks B and F with alanine abolished splicing but allowed for association. This is the first...Continue Reading

References

Mar 15, 1997·Nucleic Acids Research·F B PerlerE Adam
May 30, 1998·Nucleic Acids Research·A E Gorbalenya
Mar 26, 1998·Protein Science : a Publication of the Protein Society·S Pietrokovski
Jul 17, 1998·Current Opinion in Chemical Biology·F B PerlerH Paulus
Aug 5, 1998·Proceedings of the National Academy of Sciences of the United States of America·H WuX Q Liu
Aug 19, 1999·Protein Science : a Publication of the Protein Society·R B Kapust, D S Waugh
Nov 26, 1999·Proceedings of the National Academy of Sciences of the United States of America·C P ScottS J Benkovic
May 29, 2000·The Journal of Biological Chemistry·B W PolandF A Quiocho
Jul 13, 2000·Journal of Molecular Biology·K IchiyanagiK Morikawa
Aug 31, 2000·Annual Review of Biochemistry·H Paulus
Jan 22, 2001·The Journal of Biological Chemistry·K V Mills, H Paulus
Mar 7, 2001·Applied and Environmental Microbiology·L SunM Q Xu
Jun 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·N YadavS C Falco

❮ Previous
Next ❯

Citations

Jul 28, 2009·Journal of the American Chemical Society·Zhenming DuChunyu Wang
Jul 15, 2009·Nature Chemical Biology·Joshua A KritzerSusan Lindquist
Apr 5, 2011·Nature Structural & Molecular Biology·Brian P CallahanMarlene Belfort
May 23, 2009·Molecular BioSystems·Liyun ZhangYangzhong Liu
Nov 10, 2010·The Journal of Biological Chemistry·Liyun ZhangYangzhong Liu
May 23, 2007·Molekuliarnaia biologiia·P L Starokadomskiĭ
Feb 5, 2014·Mobile DNA·Natalya I Topilina, Kenneth V Mills
Apr 3, 2003·Proceedings of the National Academy of Sciences of the United States of America·Hang Gyeong ChinSriharsa Pradhan
Nov 14, 2014·Open Biology·Florence H GohardWilliam C Earnshaw
Aug 29, 2012·Cellular and Molecular Life Sciences : CMLS·Gerrit Volkmann, Henning D Mootz
Jun 5, 2007·FEBS Letters·Esther J PearlSigurd M Wilbanks
Dec 4, 2003·Molecular Microbiology·Jonathan CaspiShmuel Pietrokovski
Jul 27, 2011·The FEBS Journal·Gerrit Volkmann, Xiang-Qin Liu
Oct 7, 2004·Developmental Biology·Jan PielageChristian Klämbt
Mar 9, 2010·Chemistry : a European Journal·Liyun ZhangYangzhong Liu
May 5, 2007·Bioorganicheskaia khimiia·V N StepannenkoA I Miroshnikov
Dec 20, 2011·Organic & Biomolecular Chemistry·Jon I MujikaAdrian J Mulholland
Apr 4, 2014·The Journal of Biological Chemistry·Kenneth V MillsFrancine B Perler
Mar 17, 2004·The Journal of Biological Chemistry·Kenneth V MillsLisa A Wuerdeman
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
Jan 15, 2021·Biochimica Et Biophysica Acta. General Subjects·Valerio ChiariniAndrea Ilari
Mar 18, 2021·Biochimie·Sunita PandaSasmita Nayak
Mar 31, 2009·The Journal of Physical Chemistry. B·Jon I MujikaAdrian J Mulholland
Jul 25, 2017·The Journal of Physical Chemistry. B·J I Mujika, X Lopez
Aug 3, 2013·Biochemistry·Michael C NicastriKenneth V Mills
Dec 12, 2002·Chemical Reviews·Thomas C Evans, Ming-Qun Xu
Sep 26, 2021·Proteins·Leila T AlexanderTorsten Schwede

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.