Gene (Homo sapiens) | hEMC1 | NIH Mammalian Gene Collection | NCBI: BC034589 | |
Gene (Homo sapiens) | hEMC2 | NIH Mammalian Gene Collection | NCBI: BC021667 | |
Gene (Homo sapiens) | hEMC3 | NIH Mammalian Gene Collection | NCBI: BC022807 | |
Gene (Homo sapiens) | hEMC4 | Genestrand (Eurofins, Germany) | Uniprot: Q5J8M3-1 | |
Gene (Homo sapiens) | hEMC5 | NIH Mammalian Gene Collection | NCBI: BC033588 | |
Gene (Homo sapiens) | hEMC6 | NIH Mammalian Gene Collection | NCBI: BC001409 | |
Gene (Homo sapiens) | hEMC7 | NIH Mammalian Gene Collection | NCBI: BC104936 | |
Gene (Homo sapiens) | hEMC8 | NIH Mammalian Gene Collection | NCBI: BC020250 | |
Gene (Homo sapiens) | hEMC9 | NIH Mammalian Gene Collection | NCBI: BC002491 | |
Gene (Homo sapiens) | hEMC10 | Genestrand (Eurofins, Germany) | Uniprot: Q5UCC4-1 | |
Gene (Saccharomyces cerevisiae) | yEMC1 | Uniprot | Uniprot: P25574 | |
Gene (Saccharomyces cerevisiae) | yEMC2 | Uniprot | Uniprot: P47133 | |
Gene (Saccharomyces cerevisiae) | yEMC3 | Uniprot | Uniprot: P36039 | |
Gene (Saccharomyces cerevisiae) | yEMC4 | Uniprot | Uniprot: P53073 | |
Gene (Saccharomyces cerevisiae) | yEMC5 | Uniprot | Uniprot: P40540 | |
Gene (Saccharomyces cerevisiae) | yEMC6 | Uniprot | Uniprot: Q12431 | |
Gene (Saccharomyces cerevisiae) | yEMC7 | Uniprot | Uniprot: P39543 | |
Gene (Saccharomyces cerevisiae) | yEMC10 | Uniprot | Uniprot: Q12025 | |
Recombinant DNA reagent | pX458 | Addgene | pX458 | |
Recombinant DNA reagent | pKDP041 | This study; available from the Weissman Lab | Cas9-sfGFP- EMC5 sgRNA3 | single guide KO system targeting EMC5 gene |
Recombinant DNA reagent | pKDP077 | This study; available from the Weissman Lab | Cas9-sfGFP-EMC1_ sgRNA3_sgRNA4 | dual guide KO system targeting EMC1 gene |
Recombinant DNA reagent | pKDP080 | This study; available from the Weissman Lab | Cas9-sfGFP-EMC2_ sgRNA4_sgRNA5 | dual guide KO system targeting EMC2 gene |
Recombinant DNA reagent | pKDP083 | This study; available from the Weissman Lab | Cas9-sfGFP-EMC3_ sgRNA1_sgRNA2 | dual guide KO system targeting EMC3 gene |
Recombinant DNA reagent | pKDP119 | This study; available from the Weissman Lab | SFFV-insert site- IRES-Puro-P2A-BFP | parental vector |
Recombinant DNA reagent | pKDP121 | This study; available from the Weissman Lab | pTwist+Lenti+SFFV+ EMC1+IRES+Puro+ P2A+BFP+WPRE | EMC1 covering plasmid |
Recombinant DNA reagent | pKDP122 | This study; available from the Weissman Lab | pTwist+Lenti+SFFV+ EMC3+IRES+Puro+ P2A+BFP+WPRE | EMC3 covering plasmid |
Recombinant DNA reagent | pKDP124 | This study; available from the Weissman Lab | pTwist+Lenti+SFFV+ EMC5+IRES+Puro+ P2A+BFP+WPRE | EMC5 covering plasmid |
Recombinant DNA reagent | pKDP125 | This study; available from the Weissman Lab | pTwist+Lenti+SFFV+ EMC2+IRES+Puro+ P2A+BFP+WPRE | EMC2 covering plasmid |
Recombinant DNA reagent | pKDP110 | This study; available from the Weissman Lab | bAR1_mCherry_ P2A_GFP | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP111 | This study; available from the Weissman Lab | TMEM97_mCherry_ P2A_GFP | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP136 | This study; available from the Weissman Lab | GFP_P2A_mCherry_ SQS_TMD_opsintag | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_D31K | Twist; available from the Weissman Lab | hsEMC1_mut_D31K | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_R69D | Twist; available from the Weissman Lab | hsEMC1_mut_R69D | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_G71S | Twist; available from the Weissman Lab | hsEMC1_mut_G71S | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_ hsEMC1_mut_ R76D_K80D | Twist; available from the Weissman Lab | hsEMC1_mut_ R76D_K80D | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_T82M | Twist; available from the Weissman Lab | hsEMC1_mut_T82M | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_T82A | Twist; available from the Weissman Lab | hsEMC1_mut_T82A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_A144T | Twist; available from the Weissman Lab | hsEMC1_mut_A144T | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_H93D_E138D_N282K | Twist; available from the Weissman Lab | hsEMC1_mut_H93D_ E138D_N282K | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_R275E_R404E | Twist; available from the Weissman Lab | hsEMC1_mut_ R275E_R404E | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_G471R | Twist; available from the Weissman Lab | hsEMC1_mut_G471R | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_F473Y_R487K | Twist; available from the Weissman Lab | hsEMC1_mut_ F473Y_R487K | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_M483A_ R487H_Q491N | Twist; available from the Weissman Lab | hsEMC1_mut_M483A_ R487H_Q491N | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_G868R | Twist; available from the Weissman Lab | hsEMC1_mut_G868R | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_R881C | Twist; available from the Weissman Lab | hsEMC1_mut_R881C | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC1_ mut_K951A_K957A | Twist; available from the Weissman Lab | hsEMC1_mut_ K951A_K957A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_K18A_K21A | Twist; available from the Weissman Lab | hsEMC2_mut_ K18A_K21A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_R80A_R81A_ K90A_R112A | Twist; available from the Weissman Lab | hsEMC2_mut_R80A_ R81A_K90A_R112A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_K125E_R126D_K127E | Twist; available from the Weissman Lab | hsEMC2_mut_K125E_ R126D_K127E | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_N137A_N167A | Twist; available from the Weissman Lab | hsEMC2_mut_ N137A_N167A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_E146A_E149A_Q150A | Twist; available from the Weissman Lab | hsEMC2_mut_E146A_ E149A_Q150A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_E168A_D170A_K173A | Twist; available from the Weissman Lab | hsEMC2_mut_E168A_ D170A_K173A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_E206A_E209A_D252A | Twist; available from the Weissman Lab | hsEMC2_mut_E206A_ E209A_D252A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_ mut_K248E_D252K_K255E | Twist; available from the Weissman Lab | hsEMC2_mut_K248E_ D252K_K255E | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_mut_ R266A_Q269A_R273A | Twist; available from the Weissman Lab | hsEMC2_mut_R266A_ Q269A_R273A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC2_mut_ Q269A_E286A_E290A | Twist; available from the Weissman Lab | hsEMC2_mut_Q269A_ E286A_E290A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_WT | Twist; available from the Weissman Lab | hsEMC3_WT | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_D9A | Twist; available from the Weissman Lab | hsEMC3_mut_D9A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_R13E | Twist; available from the Weissman Lab | hsEMC3_mut_R13E | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_K42A_K43A | Twist; available from the Weissman Lab | hsEMC3_mut_ K42A_K43A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_E63K_D213K_E223K | Twist; available from the Weissman Lab | hsEMC3_mut_E63K_ D213K_E223K | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_K70Y | Twist; available from the Weissman Lab | hsEMC3_mut_K70Y | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_V118A_I122A | Twist; available from the Weissman Lab | hsEMC3_mut_ V118A_I122A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_N114D_N117D | Twist; available from the Weissman Lab | hsEMC3_mut_ N114D_N117D | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_R180A | Twist; available from the Weissman Lab | hsEMC3_mut_R180A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_R59E_R62E_K216E | Twist; available from the Weissman Lab | hsEMC3_mut_R59E_ R62E_K216E | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_R147E | Twist; available from the Weissman Lab | hsEMC3_mut_R147E | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_F148L | Twist; available from the Weissman Lab | hsEMC3_mut_F148L | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_M151L | Twist; available from the Weissman Lab | hsEMC3_mut_M151L | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_I186V_I182V | Twist; available from the Weissman Lab | hsEMC3_mut_ I186V_I182V | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC3_ mut_K244A_ H247A_E249A | Twist; available from the Weissman Lab | hsEMC3_mut_K244A_ H247A_E249A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_WT | Twist; available from the Weissman Lab | hsEMC5_WT | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_A18L | Twist; available from the Weissman Lab | hsEMC5_mut_A18L | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_D44K | Twist; available from the Weissman Lab | hsEMC5_mut_D44K | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_D82A_R85A | Twist; available from the Weissman Lab | hsEMC5_mut_ D82A_R85A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_F22L | Twist; available from the Weissman Lab | hsEMC5_mut_F22L | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_E75A | Twist; available from the Weissman Lab | hsEMC5_mut_E75A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_H19L_S23A_Q26L | Twist; available from the Weissman Lab | hsEMC5_mut_H19L_ S23A_Q26L | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_K7A | Twist; available from the Weissman Lab | hsEMC5_mut_K7A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_K7E | Twist; available from the Weissman Lab | hsEMC5_mut_K7E | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_R28A_R32A | Twist; available from the Weissman Lab | hsEMC5_mut_ R28A_R32A | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_I63L | Twist; available from the Weissman Lab | hsEMC5_mut_I63L | See Supplementary file 5 for sequence |
Recombinant DNA reagent | pKDP119_hsEMC5_ mut_F90A | Twist; available from the Weissman Lab | hsEMC5_mut_F90A | See Supplementary file 5 for sequence |
Antibody | Mouse GAPDH Primary Antibody | Abcam | ab8245 | See Supplementary file 5 for sequence |
Antibody | Rabbit TMEM97 primary | ThermoFisher Scientific | PA-23003 | |
Antibody | Rabbit FDFT1 Primary Antibody | Abcam | ab195046 | |
Antibody | Rat BAP31 Primary Antibody | ThermoFisher Scientific | MA3-002 | |
Antibody | Rabbit (KIAA0090) EMC1 primary antibody | Abcam | ab242112 | |
Antibody | Rabbit TTC35 (EMC2) primary antibody | Proteintech | 25443–1-AP | |
Antibody | Rabbit TM111 (EMC3) primary antibody | ThermoFisher Scientific | #711771 | |
Antibody | Rabbit EMC4 primary antibody | Abcam | ab184544 | |
Antibody | Rabbit MMGT1 (EMC5) primary antibody | Bethyl Laboratories | A305-833A-M | |
Antibody | Rabbit (C19orf63) EMC10 primary antibody | Abcam | ab180148 | |
Antibody | IRDye 800CW Goat anti-Mouse IgG Secondary Antibody | LI-COR Biosciences | 925–32210 | |
Antibody | IRDye 800CW Goat anti-Rabbit IgG Secondary Antibody | LI-COR Biosciences | 926–32211 | |
Peptide, recombinant protein | Fab DE4 | This study; available from the Weissman Lab | LMV83 | LFAIPLVVPFYSHSALDVVMTQSPLSLPV TPGEPASISCRSSQTLMNRNGNNFLDW YVQKPGQSPQLLIYLGSNRAPGVPDRFS GSGSGTDFTLKISRLEVEDVGVYYCMQA LQTPRTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC-- MAQVQLQQWGAGLLKPSETLSLTCAVYG GSFSGYYWSWIRQPPGKGLEWIGEINHS GSTNYNPSLKSRVTISVDTSKKQFSLKLS SVTAADTAVYYCARFSYYGSGIYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAA LGCLVKDYFPEPVTVSWNSGALTSGVHT FPAVLQSSGLYSLSSVVTVPSSSLGTQTYI CNVNHKPSNTKVDKKVEPKSCAAAHHH HHHGAAEQKLISEEDLNGAA- |
Peptide, recombinant protein | Fab DH4 | This study; available from the Weissman Lab | LMV82 | LFAIPLVVPFYSHSALDVVMTQSPLSLPV TPGEPASISCRSSQTLMNRNGNNFLDW YLQKPGQSPQLLIYLGSNRAPGVPDRFS GSGSGTDFTLRISRVEPEDVGVYYCMQA LQTPSFGGGTKVEIRRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGEC-- MAQVQLQQWGAGLLKPSETLSLTCAVY GGSFSGYYWSWIRQPPGKGLEWIGEIN HSGSTNYNPSLKSRVTISVDTSKNQFSL KLSSVTAADTAVYYCARGLAGRGYYGSG SYLRWGQGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTQTYICNVNHKPSNTKVDK KVEPKSCAAAHHHHHHGAAE QKLISEEDLNGAA- |
Commercial assay or kit | Superose 6, 10/300 GL | GE Healthcare | 17517201 | |
Commercial assay or kit | R1.2/1.3 200 and 300 mesh Cu holey carbon grids | Quantifoil | 1210627 | |
Commercial assay or kit | BL21 Gold Star competent cells | Invitrogen | C602003 | |
Commercial assay or kit | Anti-Flag agarose beads | Millipore | A2220 | |
Commercial assay or kit | EconoPac Chromatography Columns | Biorad | 7321010 | |
Commercial assay or kit | 100 KD MW | EMD Millipore | UFC810024 | |
Commercial assay or kit | Superose 6, 10/300 GL | Cytiva | 29-0915-96 | |
Commercial assay or kit | cOmplete EDTA-free Protease Inhibitor Cocktail | Roche | catalog No. 05056489001 | |
Commercial assay or kit | Bio-Beads | Biorad | 1523920 | |
Commercial assay or kit | R1.2/1.3 200 and 300 mesh Cu holey carbon grids | Quantifoil | 1210627 | |
Commercial assay or kit | Ultrathin Carbon Film on Lacey Carbon Support Film, 400 mesh, Copper | Ted Pella | #01824 | |
Chemical compound, drug | FuGENE HD transfection reagent | Promega | E2312 | |
Chemical compound, drug | 1-Palmitoyl-2-oleoyl-sn- glycero-3-PC (POPC) | Cayman Chemical | 15102 | |
Chemical compound, drug | Glyco-diosgenin (GDN) | Anatrace | GDN101 | |
Chemical compound, drug | yeast extract total | Avanti Polar Lipids | 190000 P-100mg | |
Chemical compound, drug | Cholesteryl Hemisuccinate Tris Salt | Anatrace | CH210 5 GM | |
Chemical compound, drug | b-DDM | Anatrace | D310 | |
Chemical compound, drug | IPTG | GoldBio | I2481C5 | |
Chemical compound, drug | EX-CELL 420 Serum-Free Medium | Sigma-Aldrich | 14420 C | |
Chemical compound, drug | FreeStyle 293 Expression Medium | Thermo fischer | 12338018 | |
Cell line (Homo sapiens) | HEK293S GnTI- | ATCC | CRL-3022 | Mycoplasma negative |
Cell line (Spodoptera frugiperda) | Sf9 | Thermo Fischer | 11496015 | |
Cell line (Homo sapiens) | K562 crispri | Gilbert et al., 2014 | K562 crispri | |
Strain, strain background Saccharomyces cerevisiae | Overexpressed EMC with yEMC5-linker- TEV-linker-3xFlag | This study; available from the Weissman Lab | LMV84 | BY4743 ---- MATa/alpha, his3∆0/his3∆0, leu2∆0/leu2∆0, LYS2/lys2∆0, met15∆0/MET15, ura3∆0/ura3∆0, emc1::NatMX:: TEF2pr-EMC1/EMC1, emc3:: KanMX::TEF2pr-EMC3/EMC3, emc4::his3(CG)::TEF2pr-EMC4/ EMC4, sop4::HphMx::TEF2pr- SOP4/SOP4, EMC2/emc2::NatMX:: TEF2pr-EMC2, emc5::EMC5-TEV- 3xFLAG::ura3(KL)/emc5::his3(CG):: TEF2pr-EMC5-TEV-3xFLAG::KanMX, EMC6/emc6::HphMX::TEF2pr-EMC6, YDR056c/ydr056c::leu2(CG):: TEF2pr-ydr056c |
Strain, strain background Saccharomyces cerevisiae | Endogenous yEMC5- linker-TEV-linker-3xFlag | This study; available from the Weissman Lab | LMV85 | W303 ---- EMC5-3xF:ura - Linker-TEV- linker-3xFlag (GGSGSGENLYFQSGSGS DYKDDDDKDYKDDDDKDYKDDDDK) |
Software, algorithm | CryoSPARC version 2.12.4. | Punjani et al., 2017 | RRID:SCR_016501 | |
Software, algorithm | UCSF ChimeraX Version 1.0 | Goddard et al., 2018 | RRID:SCR_015872 | |
Software, algorithm | PHENIX Version 1.17 | Adams et al., 2011; | RRID:SCR_014224 | |
Software, algorithm | Coot Version 0.8 | Emsley et al., 2010 | RRID:SCR_014222 | |
Software, algorithm | RELION 3.1 | Kimanius et al., 2016; Zivanov et al., 2018 | http://www2.mrclmb.cam.ac.uk/relion | |
Software, algorithm | SerialEM | Mastronarde, 2005 | RRID:SCR_017293 | |