The N-terminal methyltransferase homologs NRMT1 and NRMT2 exhibit novel regulation of activity through heterotrimer formation.

Faughn, Jonathan

doi:10.18297/etd/3012

Cited by 3 publications

(16 citation statements)

References 136 publications

(209 reference statements)

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“…The copyright holder for this preprint this version posted October 5, 2022. ; https://doi.org/10.1101/2022.10.05.510978 doi: bioRxiv preprint One form of METTL11A regulation that we have identified is through physical interaction with its homolog, METTL11B (NRMT2/NTMT2) (12). Using analytical ultracentrifugation, METTL11A was found to primarily exist as a dimer, and when combined with METTL11B, formed a heterotrimer consisting of the METTL11A dimer bound to a METTL11B monomer (12). Through this interaction, METTL11B stabilizes METTL11A and specifically promotes METTL11A methylation of non-canonical targets (12).…”

Section: Mettl11a (Nrmt1/ntmt1mentioning

confidence: 99%

“…Through this interaction, METTL11B provides stability to METTL11A and enhances METTL11A trimethylation activity on non-canonical substrates (12). Using computational modeling, we previously published a model of the METTL11A/METTL11B interaction and proposed twelve pairs of residues from METTL11A and METTL11B that are predicted to be important for mediating this interaction (12). Here, we first aimed to verify this model by mutating predicted residues and assaying the effect on METTL11A binding to METTL11B.…”

Section: Identification Of Residues That Regulate the Mettl11a/mettl1...mentioning

confidence: 99%

“…From the list of twelve predicted pairs, we prioritized mutations that also have biological relevance. Using the Catalogue of Somatic Mutations in Cancer (COSMIC), we found three METTL11B cancer-associated mutations (Q67H-lung, F68L-colon, D232N-prostate) that affected residues predicted to be important for binding from our previous model (12,35). These three mutations, and a fourth cancer-associated METTL11B mutation (V224L-breast) that is catalytically inactive but does not mediate binding to METTL11A, were all introduced individually into human METTL11B-GFP.…”

Section: Identification Of Residues That Regulate the Mettl11a/mettl1...mentioning

confidence: 99%

“…Computational models were generated with the ZDOCK server and graphics were viewed and analyzed using Chimera (36,37). This method was first used to generate a model of the METTL11A (PDB 5E1B) dimer that closely correlated with our previously published METTL11A dimer model (12). The METTL11A dimer model was then docked to a METTL11B (PDB 6DUB) monomer using the ZDOCK server, with METTL11B D232 identified as a contacting residue (Fig.…”

Section: Identification Of Residues That Regulate the Mettl11a/mettl1...mentioning

confidence: 99%

“…The METTL11A dimer model was produced using the METTL11A crystal structure of the monomer (PDB: 2EX4, chain A) as both input structures in the protein-protein docking tool, ZDOCK (36). Out of the top 10 predicted models, the one most closely resembling the previously published METTL11A dimer model (12) was selected for use in further modeling. To produce the METTL11A/METTL11B heterotrimer model with the METTL11B D232 constraint, the METTL11A dimer and the METTL11B crystal structure (PDB: 5DUB) were used as inputs in ZDOCK (36), with METTL11B D232 selected as a contacting residue.…”

Section: Molecular Modelingmentioning

confidence: 99%

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Opposing regulation of METTL11A by its family members METTL11B and METTL13

Parker

Tooley

2022

Preprint

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N-terminal protein methylation (Nα-methylation) is a post-translational modification (PTM) that influences a variety of biological processes by regulating protein stability, protein-DNA interactions, and protein-protein interactions. Although significant progress has been made in understanding the biological roles of this PTM, we still do not completely understand how the methyltransferases that place it are regulated. A common mode of methyltransferase regulation is through complex formation with close family members, and we have previously shown that the Nα-trimethylase METTL11A (NRMT1/NTMT1) is activated through binding of its close homolog METTL11B (NRMT2/NTMT2). It has also recently been reported that METTL11A co-fractionates with a third METTL family member METTL13, which methylates both the N-terminus and lysine 55 (K55) of eukaryotic elongation factor 1 alpha (eEF1A). Here we confirm a regulatory interaction between METTL11A and METTL13 and show that, while METTL11B is an activator of METTL11A, METTL13 inhibits METTL11A activity. This is the first example of a methyltransferase being opposingly regulated by different family members. Similarly, we find that METTL11A promotes the K55 methylation activity of METTL13 but inhibits its Nα-methylation activity. We also find that catalytic activity is not needed for these regulatory effects, demonstrating new, non-catalytic functions for METTL11A and METTL13. Finally, we show METTL11A, METTL11B, and METTL13 can complex together, and when all three are present, the regulatory effects of METTL13 take precedence over those of METTL11B. These findings provide a better understanding of the regulation of Nα-methylation, and suggest a model where these methyltransferases can serve in both catalytic and non-catalytic roles.

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Section: Mettl11a (Nrmt1/ntmt1mentioning

confidence: 99%

Section: Identification Of Residues That Regulate the Mettl11a/mettl1...mentioning

confidence: 99%

Section: Identification Of Residues That Regulate the Mettl11a/mettl1...mentioning

confidence: 99%

Section: Identification Of Residues That Regulate the Mettl11a/mettl1...mentioning

confidence: 99%

Section: Molecular Modelingmentioning

confidence: 99%

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Opposing regulation of METTL11A by its family members METTL11B and METTL13

Parker

Tooley

2022

Preprint

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Novel regulation of the transcription factor ZHX2 by N-terminal methylation

et al. 2022

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N-terminal methylation (Nα-methylation) by the methyltransferase NRMT1 is an important post-translational modification that regulates protein-DNA interactions.Accordingly, its loss impairs functions that are reliant on such interactions, including DNA repair and transcriptional regulation. Global loss of Nα-methylation results in severe developmental and premature aging phenotypes, but given over 300 predicted substrates, it is hard to discern which physiological substrates contribute to each phenotype. One of the most striking phenotypes in NRMT1 knockout (Nrmt1 -/-) mice is early liver degeneration. To identify the disrupted signaling pathways leading to this phenotype and the NRMT1 substrates involved, we performed RNA-sequencing analysis of control and Nrmt1 -/adult mouse livers. We found both a significant upregulation of transcripts in the cytochrome P450 (CYP) family and downregulation of transcripts in the major urinary protein (MUP) family. Interestingly, transcription of both families is inversely regulated by the transcription factor zinc fingers and homeoboxes 2 (ZHX2).ZHX2 contains a non-canonical NRMT1 consensus sequence, indicating its function could be directly regulated by Nα-methylation. We confirmed misregulation of CYP and MUP mRNA and protein levels in Nrmt1 -/livers and verified NRMT1 can methylate ZHX2 in vitro. In addition, we used mutants of ZHX2 that cannot be methylated to directly demonstrate Nα-methylation promotes ZHX2 transcription factor activity.Finally, we show Nrmt1 -/mice also exhibit early postnatal de-repression of ZHX2 targets involved in fetal liver development. Taken together, these data implicate continual ZHX2 misregulation as a driving force behind the liver phenotype seen in Nrmt1 -/mice.

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Novel regulation of the transcription factor ZHX2 by N-terminal methylation

Conner

Parker

Falcone

et al. 2021

Preprint

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N-terminal methylation (Nα-methylation) by the methyltransferase NRMT1 is an important post-translational modification that regulates protein-DNA interactions. Accordingly, its loss impairs functions that are reliant on such interactions, including DNA repair and transcriptional regulation. Global loss of Nα-methylation results in severe developmental and premature aging phenotypes, but given over 300 predicted substrates, it is hard to discern which physiological substrates contribute to each phenotype. One of the most striking phenotypes in NRMT1 knockout (Nrmt1-/-) mice is early liver degeneration. To identify the disrupted signaling pathways leading to this phenotype and the NRMT1 substrates involved, we performed RNA-sequencing analysis of control and Nrmt1-/- adult mouse livers. We found both a significant upregulation of transcripts in the cytochrome P450 (CYP) family and downregulation of transcripts in the major urinary protein (MUP) family. Interestingly, transcription of both families is inversely regulated by the transcription factor zinc fingers and homeoboxes 2 (ZHX2). ZHX2 contains a non-canonical NRMT1 consensus sequence, indicating its function could be directly regulated by Nα-methylation. We confirmed misregulation of CYP and MUP mRNA and protein levels in Nrmt1-/- livers and verified NRMT1 can methylate ZHX2 in vitro. In addition, we used mutants of ZHX2 that cannot be methylated to directly demonstrate Nα-methylation promotes ZHX2 transcription factor activity. Finally, we show Nrmt1-/- mice also exhibit early postnatal de-repression of ZHX2 targets involved in fetal liver development. Taken together, these data implicate continual ZHX2 misregulation as a driving force behind the liver phenotype seen in Nrmt1-/- mice.

show abstract

The N-terminal methyltransferase homologs NRMT1 and NRMT2 exhibit novel regulation of activity through heterotrimer formation.

Cited by 3 publications

References 136 publications

Opposing regulation of METTL11A by its family members METTL11B and METTL13

Opposing regulation of METTL11A by its family members METTL11B and METTL13

Novel regulation of the transcription factor ZHX2 by N-terminal methylation

Novel regulation of the transcription factor ZHX2 by N-terminal methylation

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