Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | METTL18 | GenBank | NM_033418 | |
| Gene (H. sapiens) | RPL3 | GenBank | NM_000967 | |
| Gene (Mus musculus) | METTL18 | GenBank | NM_027279 | cDNA clone AK139786 (FANTOM) was used |
| Gene (M. musculus) | RPL3 | GenBank | NM_013762 | |
| Cell line (H. sapiens) | Naïve HEK293T | RIKEN BRC | RCB2202 | Female |
| Cell line (H. sapiens) | METTL18 KO HEK293T | This paper | Female; CRISPR/Cas9-edited cell line, knocking out METTL18 | |
| Cell line (H. sapiens) | METTL18 KO HEK293T with stable METTL18 expression | This paper | Female; exogenous METTL18 expression was induced in METTL18 KO HEK293T | |
| Cell line (H. sapiens) | SETD3 KO HEK293T | This paper | Female; CRISPR/Cas9-edited cell line, knocking out SETD3 | |
| Cell line (H. sapiens) | SETD3-METTL18 DKO HEK293T | This paper | Female; CRISPR/Cas9-edited cell line, knocking out SETD3 and METTL18 simultaneously | |
| Cell line (H. sapiens) | Naïve HAP1 | Horizon Discovery | Cat# C631 | Male |
| Cell line (H. sapiens) | HAP1 1-nt deletion |
Horizon Discovery | Cat# HZGHC000541c009 | Male; CRISPR/Cas9-edited cell line containing a 1-nt deletion in a coding exon of METTL18 |
| Cell line (H. sapiens) | HAP1 2-nt deletion |
Horizon Discovery | Cat# HZGHC000541c012 |
Male; CRISPR/Cas9-edited cell line containing a 2-nt deletion in a coding exon of METTL18 |
| Cell line (H. sapiens) | HAP1 4-nt deletion |
Horizon Discovery | Cat# HZGHC000541c002 |
Male; CRISPR/Cas9-edited cell line containing a 4-nt deletion in a coding exon of METTL18 |
| Transfected construct (H. sapiens) | PX330-B/B-gMETTL18 | This paper | Guide RNA expression | |
| Transfected construct (H. sapiens) | pL-CRISPR.EFS.tRFP-gSETD3 | This paper | Guide RNA expression | |
| Transfected construct (H. sapiens) | pcDNA3-hRPL3-FLAG (WT and His245Ala) | This paper | Protein expression | |
| Transfected construct (H. sapiens) | pcDNA3-mRPL3-FLAG (WT and His245Ala) | This paper | Protein expression | |
| Transfected construct (H. sapiens) | pQCXIP-hMETTL18-HA | This paper | Protein expression | |
| Transfected construct (H. sapiens) | hMETTL18-Asp193Lys-Gly195Arg-Gly197Arg-HA | This paper | Protein expression | |
| Transfected construct (H. sapiens) | siRNA to RPL17 | Horizon Discovery | L-013633- 01-0005 |
|
| Transfected construct (H. sapiens) | Control siRNA | Horizon Discovery | D-001810- 10-50 |
|
| Transfected construct (H. sapiens) |
pCI-neo Fluc-EGFP | Addgene | RRID:Addgene_90170 | Protein expression |
| Transfected construct (H. sapiens) |
pCI-neo FlucDM-EGFP | Addgene | RRID:Addgene_90172 | Protein expression |
| Antibody | Anti-α-tubulin (mouse monoclonal) | Sigma-Aldrich | Cat# T5168; RRID:AB_477579 | WB 1:1000 |
| Antibody | Anti-METTL18 (rabbit polyclonal) | Proteintech Group | Cat# 25553-1-AP; RRID:AB_2503968 | WB (1:1000) |
| Antibody | Anti-SETD3 (rabbit polyclonal) | Abcam | Cat# ab174662; RRID:AB_2750852 | WB (1:1000) |
| Antibody | Anti-RPL3 (mouse monoclonal) | Proteintech Group | Cat# 66130-1-lg; RRID:AB_2881529 | WB (1:1000) |
| Antibody | Anti-RPL3 (rabbit polyclonal) | Proteintech Group | Cat# 11005-1-AP; RRID:AB_2181760 | WB (1:1000) |
| Antibody | Anti-PES1 (rat monoclonal) | Abcam | Cat# ab252849; RRID:AB_2915993 | WB (1:1000) |
| Antibody | Anti-NMD3 (rabbit monoclonal) | Abcam | Cat# ab170898; RRID:AB_2915994 | WB (1:1000) |
| Antibody | Anti-HA (mouse monoclonal) | MBL | Cat# M180-3; RRID:AB_10951811 | WB (1:1000) |
| Antibody | Anti-GFP (rabbit polyclonal) | Abcam | Cat# ab6556; RRID:AB_305564 | WB (1:1000) |
| Antibody | Anti-β-actin (mouse monoclonal) | MBL | Cat# M177-3; RRID:AB_10697039 | WB (1:1000) |
| Antibody | Anti-mouse IgG, conjugate with HRP (sheep polyclonal) | Cytiva | Cat# NA931V; RRID:AB_772210 | WB (1:5000) |
| Antibody | Anti-rabbit IgG, conjugated with HRP (donkey polyclonal) | Cytiva | Cat# NA934V; RRID:AB_772206 | WB (1:5000) |
| Antibody | Anti-mouse IgG, conjugated with IRDye 680RD (goat polyclonal) | LI-COR Biosciences | Cat# 925-68070; RRID:AB_2651128 | WB (1:10,000) |
| Antibody | Anti-rabbit IgG, conjugated with IRDye 680RD (goat polyclonal) | LI-COR Biosciences | Cat# 925-68071; RRID:AB_2721181 | WB (1:10,000) |
| Antibody | Anti-mouse IgG, conjugated with IRDye 800CW (goat polyclonal) | LI-COR Biosciences | Cat# 926-32210; RRID:AB_621842 | WB (1:10,000) |
| Antibody | Anti-rabbit IgG, conjugated with IRDye 800CW (goat polyclonal) | LI-COR Biosciences | Cat# 926-32211; RRID:AB_621843 | WB (1:10,000) |
| Antibody | Anti-rat IgG, conjugated with IRDye 800CW (goat polyclonal) | LI-COR Biosciences | Cat# 926-32219; RRID:AB_1850025 | WB (1:10,000) |
| Antibody | Anti-GFP (mouse monoclonal) | Abcam | Cat# ab1218; RRID:AB_298911 | IF (1:1000) |
| Antibody | Anti-mouse IgG, conjugated with Alexa Fluor 488 (goat polyclonal) | Thermo Fisher Scientific | Cat# R37120; RRID:AB_2556548 | IF (1:1000) |
| Recombinant DNA reagent | pET19b-mMETTL18 | This paper | Expression of N-terminally His-tagged mouse METTL18 in Escherichia coli | |
| Recombinant DNA reagent | pCold-GST-mMETTL18 | This paper | Expression of N-terminally His- and GST-tagged mouse METTL18 in E. coli | |
| Recombinant DNA reagent | Salmonella MTAN | Addgene | RRID:Addgene_64041 | Expression of Salmonella MTAN in E. coli |
| Recombinant DNA reagent | pGL3 basic | Promega | Cat# E1751 |
|
| Recombinant DNA reagent | psiCHECK2 | Promega | Cat# C8021 |
|
| Recombinant DNA reagent | psiCHECK2-Y0× | This study | Encoding Rluc-Fluc fusion | |
| Recombinant DNA reagent | psiCHECK2-Y39× | This study | Encoding Rluc-Fluc fusion with Tyr repeat insertion | |
| Sequence-based reagent | Probe for tRNATyrGUA | This paper | 5′-ACAGTCCTCCGCTCTACCAGCTGA-3′ | |
| Sequence-based reagent | Probe for tRNALeuHAG | This paper | 5′-CAGCGCCTTAGACCGCTCGGCCA-3′ | |
| Sequence-based reagent | Probe for U6 | This paper | 5′-CACGAATTTGCGTGTCATCCTT-3′ | |
| Commercial assay or kit | QuikChange Site-Directed Mutagenesis Kit | Agilent Technologies | Cat# 200518 | |
| Commercial assay or kit | PEI transfection reagent | Polysciences | ||
| Commercial assay or kit | TransIT-293 | Mirus | Cat# MIR2700 | |
| Commercial assay or kit | TransIT-X2 Dynamic Delivery System | Mirus | Cat# MIR6000 | |
| Commercial assay or kit | Dual-Luciferase Reporter Assay System | Promega | Cat# E1910 | |
| Commercial assay or kit | Rabbit Reticulocyte Lysate, Nuclease-Treated | Promega | Cat# L4960 | |
| Commercial assay or kit | Click-iT Cell Reaction Buffer Kit | Thermo Fisher Scientific | Cat# C10269 | |
| Commercial assay or kit | T7-Scribe Standard RNA IVT kit | CELLSCRIPT | Cat# C-MSC11610 | |
| Commercial assay or kit | ScriptCap m7G Capping system | CELLSCRIPT | Cat# C-SCCE0625 | |
| Commercial assay or kit | A-Plus poly(A) polymerase Tailing kit | CELLSCRIPT | Cat# C-PAP5104H | |
| Chemical compound, drug | IRdye800CW azide | LI-COR Biosciences | Cat# 929-65000 | |
| Chemical compound, drug | MG132 | FUJIFILM Wako Chemicals | Cat# 139-18451 | |
| Software, algorithm | Proteome Discoverer | Thermo Fisher Scientific | Version 2.3 | LC-MS/MS for methylated peptide |
| Software, algorithm | Proteome Discoverer | Thermo Fisher Scientific | Version 2.4 | SILAC-MS |
| Software, algorithm | MASCOT | Matrix Science | Version 2.7 | LC-MS/MS for methylated peptide and SILAC-MS |
| Software, algorithm | RELION-3.1 | https://doi.org/10.1107/S2052252520000081 | ||
| Software, algorithm | CTFFIND-4.1 | https://doi.org/10.1016/j.jsb.2015.08.008 | ||
| Software, algorithm | PHENIX | https://doi.org/10.1107/S0907444909052925 | ||
| Software, algorithm | Coot | https://doi.org/10.1107/S0907444910007493 | ||
| Software, algorithm | Image Studio | LI-COR Biosciences | Version 5.2 | |
| Software, algorithm | STAR | https://doi.org/10.1093/bioinformatics/bts635 | Version 2.7.0a | |
| Software, algorithm | kpLog | https://doi.org/10.1093/nar/gkx323 | http://kplogo.wi.mit.edu |