Cargando…

Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS

The superfamily of prokaryotic inwardly rectifying (KirBac) potassium channels is homologous to mammalian Kir channels. However, relatively little is known about their regulation or about their physiological role in vivo. In this study, we have used random mutagenesis and genetic complementation in...

Descripción completa

Detalles Bibliográficos
Autores principales: Paynter, Jennifer J., Andres-Enguix, Isabelle, Fowler, Philip W., Tottey, Stephen, Cheng, Wayland, Enkvetchakul, Decha, Bavro, Vassiliy N., Kusakabe, Yoshio, Sansom, Mark S. P., Robinson, Nigel J., Nichols, Colin G., Tucker, Stephen J.
Formato: Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3003375/
https://www.ncbi.nlm.nih.gov/pubmed/20876570
http://dx.doi.org/10.1074/jbc.M110.175687
_version_ 1782193860411129856
author Paynter, Jennifer J.
Andres-Enguix, Isabelle
Fowler, Philip W.
Tottey, Stephen
Cheng, Wayland
Enkvetchakul, Decha
Bavro, Vassiliy N.
Kusakabe, Yoshio
Sansom, Mark S. P.
Robinson, Nigel J.
Nichols, Colin G.
Tucker, Stephen J.
author_facet Paynter, Jennifer J.
Andres-Enguix, Isabelle
Fowler, Philip W.
Tottey, Stephen
Cheng, Wayland
Enkvetchakul, Decha
Bavro, Vassiliy N.
Kusakabe, Yoshio
Sansom, Mark S. P.
Robinson, Nigel J.
Nichols, Colin G.
Tucker, Stephen J.
author_sort Paynter, Jennifer J.
collection PubMed
description The superfamily of prokaryotic inwardly rectifying (KirBac) potassium channels is homologous to mammalian Kir channels. However, relatively little is known about their regulation or about their physiological role in vivo. In this study, we have used random mutagenesis and genetic complementation in K(+)-auxotrophic Escherichia coli and Saccharomyces cerevisiae to identify activatory mutations in a range of different KirBac channels. We also show that the KirBac6.1 gene (slr5078) is necessary for normal growth of the cyanobacterium Synechocystis PCC6803. Functional analysis and molecular dynamics simulations of selected activatory mutations identified regions within the slide helix, transmembrane helices, and C terminus that function as important regulators of KirBac channel activity, as well as a region close to the selectivity filter of KirBac3.1 that may have an effect on gating. In particular, the mutations identified in TM2 favor a model of KirBac channel gating in which opening of the pore at the helix-bundle crossing plays a far more important role than has recently been proposed.
format Text
id pubmed-3003375
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher American Society for Biochemistry and Molecular Biology
record_format MEDLINE/PubMed
spelling pubmed-30033752011-01-04 Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS Paynter, Jennifer J. Andres-Enguix, Isabelle Fowler, Philip W. Tottey, Stephen Cheng, Wayland Enkvetchakul, Decha Bavro, Vassiliy N. Kusakabe, Yoshio Sansom, Mark S. P. Robinson, Nigel J. Nichols, Colin G. Tucker, Stephen J. J Biol Chem Membrane Biology The superfamily of prokaryotic inwardly rectifying (KirBac) potassium channels is homologous to mammalian Kir channels. However, relatively little is known about their regulation or about their physiological role in vivo. In this study, we have used random mutagenesis and genetic complementation in K(+)-auxotrophic Escherichia coli and Saccharomyces cerevisiae to identify activatory mutations in a range of different KirBac channels. We also show that the KirBac6.1 gene (slr5078) is necessary for normal growth of the cyanobacterium Synechocystis PCC6803. Functional analysis and molecular dynamics simulations of selected activatory mutations identified regions within the slide helix, transmembrane helices, and C terminus that function as important regulators of KirBac channel activity, as well as a region close to the selectivity filter of KirBac3.1 that may have an effect on gating. In particular, the mutations identified in TM2 favor a model of KirBac channel gating in which opening of the pore at the helix-bundle crossing plays a far more important role than has recently been proposed. American Society for Biochemistry and Molecular Biology 2010-12-24 2010-09-28 /pmc/articles/PMC3003375/ /pubmed/20876570 http://dx.doi.org/10.1074/jbc.M110.175687 Text en © 2010 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) applies to Author Choice Articles
spellingShingle Membrane Biology
Paynter, Jennifer J.
Andres-Enguix, Isabelle
Fowler, Philip W.
Tottey, Stephen
Cheng, Wayland
Enkvetchakul, Decha
Bavro, Vassiliy N.
Kusakabe, Yoshio
Sansom, Mark S. P.
Robinson, Nigel J.
Nichols, Colin G.
Tucker, Stephen J.
Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS
title Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS
title_full Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS
title_fullStr Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS
title_full_unstemmed Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS
title_short Functional Complementation and Genetic Deletion Studies of KirBac Channels: ACTIVATORY MUTATIONS HIGHLIGHT GATING-SENSITIVE DOMAINS
title_sort functional complementation and genetic deletion studies of kirbac channels: activatory mutations highlight gating-sensitive domains
topic Membrane Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3003375/
https://www.ncbi.nlm.nih.gov/pubmed/20876570
http://dx.doi.org/10.1074/jbc.M110.175687
work_keys_str_mv AT paynterjenniferj functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT andresenguixisabelle functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT fowlerphilipw functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT totteystephen functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT chengwayland functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT enkvetchakuldecha functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT bavrovassiliyn functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT kusakabeyoshio functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT sansommarksp functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT robinsonnigelj functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT nicholscoling functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains
AT tuckerstephenj functionalcomplementationandgeneticdeletionstudiesofkirbacchannelsactivatorymutationshighlightgatingsensitivedomains