Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction

The methyltransferase FliB posttranslationally modifies surface-exposed ɛ-N-lysine residues of flagellin, the protomer of the flagellar filament in Salmonella enterica (S. enterica). Flagellin methylation, reported originally in 1959, was recently shown to enhance host cell adhesion and invasion by...

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Autores principales:	Chu Wang, Christian Nehls, Dirk Baabe, Olaf Burghaus, Robert Hurwitz, Thomas Gutsmann, Martin Bröring, Michael Kolbe
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/26208fccf0c6414481c713bbf711acd0
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spelling	oai:doaj.org-article:26208fccf0c6414481c713bbf711acd02021-11-25T05:47:33ZFlagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction1553-73661553-7374https://doaj.org/article/26208fccf0c6414481c713bbf711acd02021-11-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8598068/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374The methyltransferase FliB posttranslationally modifies surface-exposed ɛ-N-lysine residues of flagellin, the protomer of the flagellar filament in Salmonella enterica (S. enterica). Flagellin methylation, reported originally in 1959, was recently shown to enhance host cell adhesion and invasion by increasing the flagellar hydrophobicity. The role of FliB in this process, however, remained enigmatic. In this study, we investigated the properties and mechanisms of FliB from S. enterica in vivo and in vitro. We show that FliB is an S-adenosylmethionine (SAM) dependent methyltransferase, forming a membrane associated oligomer that modifies flagellin in the bacterial cytosol. Using X-band electron paramagnetic resonance (EPR) spectroscopy, zero-field 57Fe Mössbauer spectroscopy, methylation assays and chromatography coupled mass spectrometry (MS) analysis, we further found that FliB contains an oxygen sensitive [4Fe-4S] cluster that is essential for the methyl transfer reaction and might mediate a radical mechanism. Our data indicate that the [4Fe-4S] cluster is coordinated by a cysteine rich motif in FliB that is highly conserved among multiple genera of the Enterobacteriaceae family. Author summary The bacterial flagella are tail-like appendages that play important roles in motility and host cell infection. In Salmonella, the surface of the flagellar filaments is heavily modified by methylation on their lysine residues, contributing to efficient gut colonization and successful invasion of the host. However, little is known about the properties of the methylase FliB, that catalyzes flagellar methylation. In this study, we isolated and characterized FliB of S. enterica for the first time. We show that FliB forms oligomers that localize in close proximity to the bacterial cell membrane. The FliB methyltransferase coordinates a [4Fe-4S] cluster at its active site by several cysteine residues that were highly conserved in many entero-pathogens. We further found that FliB modifies flagellar subunits using an iron-sulfur cluster dependent mechanism and requires a reducing environment. Our work sets a stage for understanding the roles of oxygen sensitive iron-sulfur cluster in flagellar modification and entero-bacterial pathogenesis.Chu WangChristian NehlsDirk BaabeOlaf BurghausRobert HurwitzThomas GutsmannMartin BröringMichael KolbePublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 17, Iss 11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Chu Wang Christian Nehls Dirk Baabe Olaf Burghaus Robert Hurwitz Thomas Gutsmann Martin Bröring Michael Kolbe Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction
description	The methyltransferase FliB posttranslationally modifies surface-exposed ɛ-N-lysine residues of flagellin, the protomer of the flagellar filament in Salmonella enterica (S. enterica). Flagellin methylation, reported originally in 1959, was recently shown to enhance host cell adhesion and invasion by increasing the flagellar hydrophobicity. The role of FliB in this process, however, remained enigmatic. In this study, we investigated the properties and mechanisms of FliB from S. enterica in vivo and in vitro. We show that FliB is an S-adenosylmethionine (SAM) dependent methyltransferase, forming a membrane associated oligomer that modifies flagellin in the bacterial cytosol. Using X-band electron paramagnetic resonance (EPR) spectroscopy, zero-field 57Fe Mössbauer spectroscopy, methylation assays and chromatography coupled mass spectrometry (MS) analysis, we further found that FliB contains an oxygen sensitive [4Fe-4S] cluster that is essential for the methyl transfer reaction and might mediate a radical mechanism. Our data indicate that the [4Fe-4S] cluster is coordinated by a cysteine rich motif in FliB that is highly conserved among multiple genera of the Enterobacteriaceae family. Author summary The bacterial flagella are tail-like appendages that play important roles in motility and host cell infection. In Salmonella, the surface of the flagellar filaments is heavily modified by methylation on their lysine residues, contributing to efficient gut colonization and successful invasion of the host. However, little is known about the properties of the methylase FliB, that catalyzes flagellar methylation. In this study, we isolated and characterized FliB of S. enterica for the first time. We show that FliB forms oligomers that localize in close proximity to the bacterial cell membrane. The FliB methyltransferase coordinates a [4Fe-4S] cluster at its active site by several cysteine residues that were highly conserved in many entero-pathogens. We further found that FliB modifies flagellar subunits using an iron-sulfur cluster dependent mechanism and requires a reducing environment. Our work sets a stage for understanding the roles of oxygen sensitive iron-sulfur cluster in flagellar modification and entero-bacterial pathogenesis.
format	article
author	Chu Wang Christian Nehls Dirk Baabe Olaf Burghaus Robert Hurwitz Thomas Gutsmann Martin Bröring Michael Kolbe
author_facet	Chu Wang Christian Nehls Dirk Baabe Olaf Burghaus Robert Hurwitz Thomas Gutsmann Martin Bröring Michael Kolbe
author_sort	Chu Wang
title	Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction
title_short	Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction
title_full	Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction
title_fullStr	Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction
title_full_unstemmed	Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction
title_sort	flagellin lysine methyltransferase flib catalyzes a [4fe-4s] mediated methyl transfer reaction
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/26208fccf0c6414481c713bbf711acd0
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Flagellin lysine methyltransferase FliB catalyzes a [4Fe-4S] mediated methyl transfer reaction

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