Multimethylation of Rickettsia OmpB Catalyzed by Lysine Methyltransferases

Abeykoon, A.; Wang, Guanghui; Chao, Chien‐Chung; Chock, P. Boon; Guček, Marjan; Ching, Wei‐Mei; Yang, David C.H.

doi:10.1074/jbc.m113.535567

Cited by 25 publications

(26 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…rickettsii is an obligate intracellular pathogen which mainly parasites vascular endothelial cells. OmpB is the most abundant and high-molecular-mass surface protein [27] and an important virulence factor [28] of rickettsiae [29]. Adr2, a rickettsial adhesion, is also abundant on the surface of rickettsiae [30].…”

Section: Discussionmentioning

confidence: 99%

Enhanced protection against Rickettsia rickettsii infection in C3H/HeN mice by immunization with a combination of a recombinant adhesin rAdr2 and a protein fragment rOmpB-4 derived from outer membrane protein B

Gong

Wang

Xiong

et al. 2015

Vaccine

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Enhanced protection against Rickettsia rickettsii infection in C3H/HeN mice by immunization with a combination of a recombinant adhesin rAdr2 and a protein fragment rOmpB-4 derived from outer membrane protein B

Gong

Wang

Xiong

et al. 2015

Vaccine

View full text Add to dashboard Cite

“…the bacterial PrmA and PKMT1/PKMT2 as well as human METTL20, have been shown to methylate several Lys residues within the same substrate (13,14,23,43). However, AtMETTL20 is the first 7BS KMT shown to methylate two different substrates, and thus it possesses unprecedented dual substrate specificity.…”

Section: Discussionmentioning

confidence: 99%

“…Of these, PrmA, which targets ribosomal protein RpL11, has been studied in the most detail and appears to be present in all bacteria (36). Pseudomonas and certain other species of ␥-proteobacteria and firmicutes possess a 7BS MTase, EftM (EF-Tu-modifying enzyme), which methylates Lys-5 in translation elongation factor EF-Tu, thus promoting bacterial infectivity (42) Similarly, two related 7BS MTases, PKMT1 and PKMT2, that methylate the outer membrane protein OmpB on several Lys residues in Rickettsia subspecies, are also important for bacterial virulence (43,44). Thus, bacterial KMTs seem to primarily target components of the translational machinery as well as determinants of bacterial virulence.…”

Section: Discussionmentioning

confidence: 99%

The METTL20 Homologue from Agrobacterium tumefaciens Is a Dual Specificity Protein-lysine Methyltransferase That Targets Ribosomal Protein L7/L12 and the β Subunit of Electron Transfer Flavoprotein (ETFβ)

Małecki

Dahl

Moen

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Human METTL20 is a mitochondrial, lysine-specific methyltransferase that methylates the ␤-subunit of electron transfer flavoprotein (ETF␤). Interestingly, putative METTL20 orthologues are found in a subset of ␣-proteobacteria, including Agrobacterium tumefaciens. Using an activity-based approach, we identified in bacterial extracts two substrates of recombinant METTL20 from A. tumefaciens (AtMETTL20), namely ETF␤ and the ribosomal protein RpL7/L12. We show that AtMETTL20, analogous to the human enzyme, methylates ETF␤ on Lys-193 and Lys-196 both in vitro and in vivo. ETF plays a key role in mediating electron transfer from various dehydrogenases, and we found that its electron transferring ability was diminished by AtMETTL20-mediated methylation of ETF␤. Somewhat surprisingly, AtMETTL20 also catalyzed monomethylation of RpL7/L12 on Lys-86, a common modification also found in many bacteria that lack METTL20. Thus, we here identify AtMETTL20 as the first enzyme catalyzing RpL7/ L12 methylation. In summary, here we have identified and characterized a novel bacterial lysine-specific methyltransferase with unprecedented dual substrate specificity within the seven ␤-strand class of lysine-specific methyltransferases, as it targets two apparently unrelated substrates, ETF␤ and RpL7/L12. Moreover, the present work establishes METTL20-mediated methylation of ETF␤ as the first lysine methylation event occurring in both bacteria and humans.

show abstract

“…Despite recent discoveries of extensive methylation in bacterial proteins, including lysine methylations, and their roles in bacterial physiology and infection (4,7,19,20), there still exists a significant gap in our mechanistic understanding of bacterial lysine methyltransferases. Among bacterial lysine methyltransferases, the majority of class I Rossman fold methyltransferases recognize the globular body of their protein substrate and possess a shallow and relatively surface exposed catalytic center.…”

Section: Discussionmentioning

confidence: 99%

“…The human protein SUV39H1, for example, methylates the crucial surface-exposed mycobacterial protein HupB which reduces bacterial adhesion and survival inside the host cell (6). In rickettsia, methylation of outer membrane protein B (OmpB) directly controls virulence (7): virulent species display lysine trimethylation of OmpB, which mediates host adhesion, attachment, and invasion, whereas avirulent strains possess monomethylated OmpB. Finally, our previous work has revealed a role in establishment of P. aeruginosa infection for the trimethylation of translation elongation factor thermo-unstable (EF-Tu) on its Lys5 residue by the EF-Tu methyltransferase, EftM (8)(9)(10).…”

mentioning

confidence: 99%

Substrate recognition by the Pseudomonas aeruginosa EF-Tu methyltransferase EftM

Kuiper

Dey

LaMore

et al. 2019

Preprint

View full text Add to dashboard Cite

Pseudomonas aeruginosa is an opportunistic pathogen and a leading cause of serious infections in individuals with cystic fibrosis, compromised immune systems, and severe burns. During infection, P. aeruginosa adhesion to host epithelial cells is enhanced by surface exposed translation elongation factor EF-Tu carrying a Lys5 trimethylation. This modification is incorporated by the S-adenosyl-Lmethionine-dependent methyltransferase EftM. Thus, EF-Tu modification by EftM may represent a novel target to restrict the establishment of P. aeruginosa infections in vulnerable individuals. Here, we extend our understanding of EftM action by defining the molecular mechanism of EF-Tu substrate recognition by this enzyme. First, following the observation that EftM can bind to EF-Tu lacking an N-terminal peptide (encompassing the Lys5 target site), an EftM homology model was generated and used in protein-protein docking studies to predict EftM:EF-Tu interactions. The predicted protein-protein interface was then experimentally validated using site-directed mutagenesis of residues in both proteins coupled with binding and methyltransferase activity assays. We also show that EftM is unable to methylate the isolated N-terminal EF-Tu peptide and that binding-induced conformational changes in EftM are likely needed to allow placement of the first 5-6 amino acids of EF-Tu into the conserved peptide binding channel. In this channel, a

show abstract

Multimethylation of Rickettsia OmpB Catalyzed by Lysine Methyltransferases

Cited by 25 publications

References 39 publications

Enhanced protection against Rickettsia rickettsii infection in C3H/HeN mice by immunization with a combination of a recombinant adhesin rAdr2 and a protein fragment rOmpB-4 derived from outer membrane protein B

Enhanced protection against Rickettsia rickettsii infection in C3H/HeN mice by immunization with a combination of a recombinant adhesin rAdr2 and a protein fragment rOmpB-4 derived from outer membrane protein B

The METTL20 Homologue from Agrobacterium tumefaciens Is a Dual Specificity Protein-lysine Methyltransferase That Targets Ribosomal Protein L7/L12 and the β Subunit of Electron Transfer Flavoprotein (ETFβ)

Substrate recognition by the Pseudomonas aeruginosa EF-Tu methyltransferase EftM

Contact Info

Product

Resources

About