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Sequential formation of two branched intermediates during protein splicing of class three inteins

Inteins are the protein equivalent of introns. They are seamlessly removed during post-translational maturation of their host protein (extein). Inteins from extremophiles played a key role in understanding intein-mediated protein splicing. There are currently three classes of inteins defined by cata...

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Autores principales: Tori, Kazuo, Perler, Francine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Japan 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5222942/
https://www.ncbi.nlm.nih.gov/pubmed/27704298
http://dx.doi.org/10.1007/s00792-016-0876-0
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author Tori, Kazuo
Perler, Francine
author_facet Tori, Kazuo
Perler, Francine
author_sort Tori, Kazuo
collection PubMed
description Inteins are the protein equivalent of introns. They are seamlessly removed during post-translational maturation of their host protein (extein). Inteins from extremophiles played a key role in understanding intein-mediated protein splicing. There are currently three classes of inteins defined by catalytic mechanism and sequence signatures. This study demonstrates splicing of three class 3 mini-inteins: Burkholderia vietnamiensis G4 Bvi IcmO intein, Mycobacterium smegmatis MC2 155 Msm DnaB-1 intein and Mycobacterium leprae strain TN Mle DnaB intein. B. vietnamiensis has a broad ecological range and remediates trichloroethene. M. smegmatis is a biofilm forming soil bacteria. Although other intein classes have only a single branched intermediate at the C-terminal splice junction, the class 3 intein reaction pathway includes two branched intermediates. The class 3 specific branched intermediate is formed by an internal cysteine, while the C-terminal branch intermediate is at a serine or threonine in all class 3 inteins except the Bvi IcmO intein, where it is a cysteine. This latter cysteine was unable to compensate for mutation of the class 3-specific internal catalytic cysteine despite the Bvi IcmO intein having an N-terminal splice junction naturally tuned for a cysteine nucleophile, demonstrating the mandatory order of branch intermediates in class 3 inteins.
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spelling pubmed-52229422017-01-19 Sequential formation of two branched intermediates during protein splicing of class three inteins Tori, Kazuo Perler, Francine Extremophiles Special Feature: Original Paper Inteins are the protein equivalent of introns. They are seamlessly removed during post-translational maturation of their host protein (extein). Inteins from extremophiles played a key role in understanding intein-mediated protein splicing. There are currently three classes of inteins defined by catalytic mechanism and sequence signatures. This study demonstrates splicing of three class 3 mini-inteins: Burkholderia vietnamiensis G4 Bvi IcmO intein, Mycobacterium smegmatis MC2 155 Msm DnaB-1 intein and Mycobacterium leprae strain TN Mle DnaB intein. B. vietnamiensis has a broad ecological range and remediates trichloroethene. M. smegmatis is a biofilm forming soil bacteria. Although other intein classes have only a single branched intermediate at the C-terminal splice junction, the class 3 intein reaction pathway includes two branched intermediates. The class 3 specific branched intermediate is formed by an internal cysteine, while the C-terminal branch intermediate is at a serine or threonine in all class 3 inteins except the Bvi IcmO intein, where it is a cysteine. This latter cysteine was unable to compensate for mutation of the class 3-specific internal catalytic cysteine despite the Bvi IcmO intein having an N-terminal splice junction naturally tuned for a cysteine nucleophile, demonstrating the mandatory order of branch intermediates in class 3 inteins. Springer Japan 2016-10-04 2017 /pmc/articles/PMC5222942/ /pubmed/27704298 http://dx.doi.org/10.1007/s00792-016-0876-0 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Special Feature: Original Paper
Tori, Kazuo
Perler, Francine
Sequential formation of two branched intermediates during protein splicing of class three inteins
title Sequential formation of two branched intermediates during protein splicing of class three inteins
title_full Sequential formation of two branched intermediates during protein splicing of class three inteins
title_fullStr Sequential formation of two branched intermediates during protein splicing of class three inteins
title_full_unstemmed Sequential formation of two branched intermediates during protein splicing of class three inteins
title_short Sequential formation of two branched intermediates during protein splicing of class three inteins
title_sort sequential formation of two branched intermediates during protein splicing of class three inteins
topic Special Feature: Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5222942/
https://www.ncbi.nlm.nih.gov/pubmed/27704298
http://dx.doi.org/10.1007/s00792-016-0876-0
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