The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin

ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin. Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Monica Rolando [verfasserIn] Ivy Yeuk Wah Chung [verfasserIn] Caishuang Xu [verfasserIn] Laura Gomez-Valero [verfasserIn] Patrick England [verfasserIn] Miroslaw Cygler [verfasserIn] Carmen Buchrieser [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2023

Schlagwörter:	Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14

Übergeordnetes Werk:	In: mBio - American Society for Microbiology, 2010, 14(2023), 5
Übergeordnetes Werk:	volume:14 ; year:2023 ; number:5

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1128/mbio.01655-23

Katalog-ID:	DOAJ090976487

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520			\|a ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin.
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allfields	10.1128/mbio.01655-23 doi (DE-627)DOAJ090976487 (DE-599)DOAJ26ec4e0656494b79ac181002d73bfbb5 DE-627 ger DE-627 rakwb eng QR1-502 Monica Rolando verfasserin aut The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin. Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14 Microbiology Ivy Yeuk Wah Chung verfasserin aut Caishuang Xu verfasserin aut Laura Gomez-Valero verfasserin aut Patrick England verfasserin aut Miroslaw Cygler verfasserin aut Carmen Buchrieser verfasserin aut In mBio American Society for Microbiology, 2010 14(2023), 5 (DE-627)627613543 (DE-600)2557172-2 21507511 nnns volume:14 year:2023 number:5 https://doi.org/10.1128/mbio.01655-23 kostenfrei https://doaj.org/article/26ec4e0656494b79ac181002d73bfbb5 kostenfrei https://journals.asm.org/doi/10.1128/mbio.01655-23 kostenfrei https://doaj.org/toc/2150-7511 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2023 5
spelling	10.1128/mbio.01655-23 doi (DE-627)DOAJ090976487 (DE-599)DOAJ26ec4e0656494b79ac181002d73bfbb5 DE-627 ger DE-627 rakwb eng QR1-502 Monica Rolando verfasserin aut The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin. Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14 Microbiology Ivy Yeuk Wah Chung verfasserin aut Caishuang Xu verfasserin aut Laura Gomez-Valero verfasserin aut Patrick England verfasserin aut Miroslaw Cygler verfasserin aut Carmen Buchrieser verfasserin aut In mBio American Society for Microbiology, 2010 14(2023), 5 (DE-627)627613543 (DE-600)2557172-2 21507511 nnns volume:14 year:2023 number:5 https://doi.org/10.1128/mbio.01655-23 kostenfrei https://doaj.org/article/26ec4e0656494b79ac181002d73bfbb5 kostenfrei https://journals.asm.org/doi/10.1128/mbio.01655-23 kostenfrei https://doaj.org/toc/2150-7511 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2023 5
allfields_unstemmed	10.1128/mbio.01655-23 doi (DE-627)DOAJ090976487 (DE-599)DOAJ26ec4e0656494b79ac181002d73bfbb5 DE-627 ger DE-627 rakwb eng QR1-502 Monica Rolando verfasserin aut The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin. Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14 Microbiology Ivy Yeuk Wah Chung verfasserin aut Caishuang Xu verfasserin aut Laura Gomez-Valero verfasserin aut Patrick England verfasserin aut Miroslaw Cygler verfasserin aut Carmen Buchrieser verfasserin aut In mBio American Society for Microbiology, 2010 14(2023), 5 (DE-627)627613543 (DE-600)2557172-2 21507511 nnns volume:14 year:2023 number:5 https://doi.org/10.1128/mbio.01655-23 kostenfrei https://doaj.org/article/26ec4e0656494b79ac181002d73bfbb5 kostenfrei https://journals.asm.org/doi/10.1128/mbio.01655-23 kostenfrei https://doaj.org/toc/2150-7511 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2023 5
allfieldsGer	10.1128/mbio.01655-23 doi (DE-627)DOAJ090976487 (DE-599)DOAJ26ec4e0656494b79ac181002d73bfbb5 DE-627 ger DE-627 rakwb eng QR1-502 Monica Rolando verfasserin aut The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin. Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14 Microbiology Ivy Yeuk Wah Chung verfasserin aut Caishuang Xu verfasserin aut Laura Gomez-Valero verfasserin aut Patrick England verfasserin aut Miroslaw Cygler verfasserin aut Carmen Buchrieser verfasserin aut In mBio American Society for Microbiology, 2010 14(2023), 5 (DE-627)627613543 (DE-600)2557172-2 21507511 nnns volume:14 year:2023 number:5 https://doi.org/10.1128/mbio.01655-23 kostenfrei https://doaj.org/article/26ec4e0656494b79ac181002d73bfbb5 kostenfrei https://journals.asm.org/doi/10.1128/mbio.01655-23 kostenfrei https://doaj.org/toc/2150-7511 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2023 5
allfieldsSound	10.1128/mbio.01655-23 doi (DE-627)DOAJ090976487 (DE-599)DOAJ26ec4e0656494b79ac181002d73bfbb5 DE-627 ger DE-627 rakwb eng QR1-502 Monica Rolando verfasserin aut The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin 2023 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin. Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14 Microbiology Ivy Yeuk Wah Chung verfasserin aut Caishuang Xu verfasserin aut Laura Gomez-Valero verfasserin aut Patrick England verfasserin aut Miroslaw Cygler verfasserin aut Carmen Buchrieser verfasserin aut In mBio American Society for Microbiology, 2010 14(2023), 5 (DE-627)627613543 (DE-600)2557172-2 21507511 nnns volume:14 year:2023 number:5 https://doi.org/10.1128/mbio.01655-23 kostenfrei https://doaj.org/article/26ec4e0656494b79ac181002d73bfbb5 kostenfrei https://journals.asm.org/doi/10.1128/mbio.01655-23 kostenfrei https://doaj.org/toc/2150-7511 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2023 5
language	English
source	In mBio 14(2023), 5 volume:14 year:2023 number:5
sourceStr	In mBio 14(2023), 5 volume:14 year:2023 number:5
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14 Microbiology
isfreeaccess_bool	true
container_title	mBio
authorswithroles_txt_mv	Monica Rolando @@aut@@ Ivy Yeuk Wah Chung @@aut@@ Caishuang Xu @@aut@@ Laura Gomez-Valero @@aut@@ Patrick England @@aut@@ Miroslaw Cygler @@aut@@ Carmen Buchrieser @@aut@@
publishDateDaySort_date	2023-01-01T00:00:00Z
hierarchy_top_id	627613543
id	DOAJ090976487
language_de	englisch
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callnumber-first	Q - Science
author	Monica Rolando
spellingShingle	Monica Rolando misc QR1-502 misc Legionella pneumophila misc host-pathogen interactions misc histone methyltransferase misc epigenetics misc H3K14 misc Microbiology The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin
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topic_title	QR1-502 The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin Legionella pneumophila host-pathogen interactions histone methyltransferase epigenetics H3K14
topic	misc QR1-502 misc Legionella pneumophila misc host-pathogen interactions misc histone methyltransferase misc epigenetics misc H3K14 misc Microbiology
topic_unstemmed	misc QR1-502 misc Legionella pneumophila misc host-pathogen interactions misc histone methyltransferase misc epigenetics misc H3K14 misc Microbiology
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title	The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin
ctrlnum	(DE-627)DOAJ090976487 (DE-599)DOAJ26ec4e0656494b79ac181002d73bfbb5
title_full	The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin
author_sort	Monica Rolando
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author_browse	Monica Rolando Ivy Yeuk Wah Chung Caishuang Xu Laura Gomez-Valero Patrick England Miroslaw Cygler Carmen Buchrieser
container_volume	14
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author-letter	Monica Rolando
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title_sort	set and ankyrin domains of the secreted legionella pneumophila histone methyltransferase work together to modify host chromatin
callnumber	QR1-502
title_auth	The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin
abstract	ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin.
abstractGer	ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin.
abstract_unstemmed	ABSTRACT Legionella pneumophila is a bacterial pathogen ubiquitous in natural and man-made aquatic environments, where it replicates in protozoa. Its intracellular life cycle depends on the establishment of a Legionella-containing vacuole (LCV) where the bacteria replicate. To form the LCV, L. pneumophila depends on a type-4 secretion system that secretes more than 300 proteins in the host cell. Among these is RomA that encodes an N-terminal catalytic methyltransferase domain with a SET fold and a C-terminal domain containing six ankyrin repeats. We have reported that the SET domain has histone methyltransferase activity and specifically methylates lysine 14 of histone H3 during infection; however, the role of the ankyrin repeats is unknown. Here, we report the structure of RomA co-crystallized with a 15-mer peptide corresponding to the N-terminal tail of histone H3 at 2.2 Å. This structure revealed that the N-terminal arm of the peptide from Thr3-Ala7 binds to the ankyrin repeat domain and the C-terminal arm from Gly13-Arg17 and interacts with the SET domain, inserting Lys14 into the active site for methylation. Importantly, the interactions of the five N-terminal residues of the H3-peptide with the ankyrin domains account for ~50% of the peptide-RomA contacts, predicting that the ankyrin domains of RomA are essential for histone H3 binding and subsequent lysine methylation. In vitro enzymatic activity and binding assays confirmed these predictions. Thus, the six ankyrin domains are essential for chromatin binding, thereby allowing the SET domain to fulfill its histone transferase activity and to modify the host chromatin to the advantage of the pathogen. IMPORTANCE Legionella pneumophila is an intracellular bacterium responsible of Legionnaires’ disease, a severe pneumonia that is often fatal when not treated promptly. The pathogen’s ability to efficiently colonize the host resides in its ability to replicate intracellularly. Essential for intracellular replication is translocation of many different protein effectors via a specialized secretion system. One of them, called RomA, binds and directly modifies the host chromatin at a unique site (tri-methylation of lysine 14 of histone H3 [H3K14me]). However, the molecular mechanisms of binding are not known. Here, we resolve this question through structural characterization of RomA together with the H3 peptide. We specifically reveal an active role of the ankyrin repeats located in its C-terminal in the interaction with the histone H3 tail. Indeed, without the ankyrin domains, RomA loses its ability to act as histone methyltransferase. These results discover the molecular mechanisms by which a bacterial histone methyltransferase that is conserved in L. pneumophila strains acts to modify chromatin.
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container_issue	5
title_short	The SET and ankyrin domains of the secreted Legionella pneumophila histone methyltransferase work together to modify host chromatin
url	https://doi.org/10.1128/mbio.01655-23 https://doaj.org/article/26ec4e0656494b79ac181002d73bfbb5 https://journals.asm.org/doi/10.1128/mbio.01655-23 https://doaj.org/toc/2150-7511
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author2	Ivy Yeuk Wah Chung Caishuang Xu Laura Gomez-Valero Patrick England Miroslaw Cygler Carmen Buchrieser
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