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- EMDB-41300: Inner spoke ring of the yeast NPC -

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Basic information

Entry
Database: EMDB / ID: EMD-41300
TitleInner spoke ring of the yeast NPC
Map datac8 inner spoke ring recombined from multibody
Sample
  • Complex: Nuclear Pore Complex
    • Protein or peptide: x 11 types
Keywordsnuclear pore complex / nucleocytoplasmic transport / nucleoporin / membrane protein / translocase / TRANSPORT PROTEIN
Function / homology
Function and homology information


response to spindle checkpoint signaling / nuclear pore linkers / regulation of protein desumoylation / mRNA export from nucleus in response to heat stress / nuclear pore inner ring / protein localization to nuclear inner membrane / chromosome, subtelomeric region / transcription-dependent tethering of RNA polymerase II gene DNA at nuclear periphery / nuclear pore central transport channel / regulation of nucleocytoplasmic transport ...response to spindle checkpoint signaling / nuclear pore linkers / regulation of protein desumoylation / mRNA export from nucleus in response to heat stress / nuclear pore inner ring / protein localization to nuclear inner membrane / chromosome, subtelomeric region / transcription-dependent tethering of RNA polymerase II gene DNA at nuclear periphery / nuclear pore central transport channel / regulation of nucleocytoplasmic transport / nuclear pore organization / nuclear pore complex assembly / telomere tethering at nuclear periphery / post-transcriptional tethering of RNA polymerase II gene DNA at nuclear periphery / nuclear pore cytoplasmic filaments / Transport of Mature mRNA derived from an Intron-Containing Transcript / Regulation of HSF1-mediated heat shock response / nuclear pore nuclear basket / tRNA export from nucleus / SUMOylation of SUMOylation proteins / protein localization to kinetochore / SUMOylation of RNA binding proteins / structural constituent of nuclear pore / SUMOylation of chromatin organization proteins / RNA export from nucleus / nucleocytoplasmic transport / poly(A)+ mRNA export from nucleus / nuclear localization sequence binding / regulation of mitotic nuclear division / NLS-bearing protein import into nucleus / ribosomal large subunit export from nucleus / mRNA transport / nuclear pore / ribosomal small subunit export from nucleus / nuclear periphery / chromosome segregation / promoter-specific chromatin binding / molecular condensate scaffold activity / heterochromatin formation / phospholipid binding / protein import into nucleus / protein transport / nuclear envelope / single-stranded DNA binding / nuclear membrane / amyloid fibril formation / cell division / chromatin binding / protein-containing complex binding / positive regulation of DNA-templated transcription / DNA binding / RNA binding / identical protein binding / nucleus
Similarity search - Function
RNA-recognition motif (RRM) Nup35-type domain / Nucleoporin, NUP53 / Nup53/35/40-type RNA recognition motif / RNA-recognition motif (RRM) Nup35-type domain profile. / Nuclear pore protein Nup188, C-terminal / Nuclear pore protein NUP188 C-terminal domain / Nucleoporin Nup188, N-terminal / Nucleoporin Nup188, N-terminal / Nucleoporin Nup54/Nup57/Nup44 / Nucleoporin p58/p45 ...RNA-recognition motif (RRM) Nup35-type domain / Nucleoporin, NUP53 / Nup53/35/40-type RNA recognition motif / RNA-recognition motif (RRM) Nup35-type domain profile. / Nuclear pore protein Nup188, C-terminal / Nuclear pore protein NUP188 C-terminal domain / Nucleoporin Nup188, N-terminal / Nucleoporin Nup188, N-terminal / Nucleoporin Nup54/Nup57/Nup44 / Nucleoporin p58/p45 / Nucleoporin Nup54, alpha-helical domain / Nucleoporin Nup188 / : / Nucleoporin complex subunit 54 / Nucleoporin Nup188, N-terminal subdomain III / Nucleoporin, NSP1-like, C-terminal / Nucleoporin NSP1/NUP62 / Nsp1-like C-terminal region / Nucleoporin, Nup155-like / Nucleoporin, Nup155-like, C-terminal, subdomain 1 / Nucleoporin, Nup155-like, C-terminal, subdomain 2 / Nucleoporin Nup186/Nup192/Nup205 / Nuclear pore complex scaffold, nucleoporins 186/192/205 / Nucleoporin, Nup133/Nup155-like, C-terminal / Non-repetitive/WGA-negative nucleoporin C-terminal / Nucleoporin interacting component Nup93/Nic96 / Nup93/Nic96 / Nucleoporin FG repeat / Nucleoporin FG repeat region / Nucleoporin, Nup133/Nup155-like, N-terminal / Nup133 N terminal like / RNA-binding domain superfamily / Nucleotide-binding alpha-beta plait domain superfamily
Similarity search - Domain/homology
Nucleoporin NSP1 / Nucleoporin NIC96 / Nucleoporin NUP170 / Nucleoporin NUP157 / Nucleoporin NUP192 / Nucleoporin NUP57 / Nucleoporin NUP188 / Nucleoporin NUP49/NSP49 / Nucleoporin NUP53 / Nucleoporin ASM4
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 6.6 Å
AuthorsAkey CW / Echeverria I / Ouch C / Fernandez-Martinez J / Rout MP
Funding support United States, 4 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)NIH R01 GM45377 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)NIH P41 GM109824 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)NIH R01 GM112108 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)NIH GM117212 United States
CitationJournal: Mol Cell / Year: 2023
Title: Implications of a multiscale structure of the yeast nuclear pore complex.
Authors: Christopher W Akey / Ignacia Echeverria / Christna Ouch / Ilona Nudelman / Yi Shi / Junjie Wang / Brian T Chait / Andrej Sali / Javier Fernandez-Martinez / Michael P Rout /
Abstract: Nuclear pore complexes (NPCs) direct the nucleocytoplasmic transport of macromolecules. Here, we provide a composite multiscale structure of the yeast NPC, based on improved 3D density maps from ...Nuclear pore complexes (NPCs) direct the nucleocytoplasmic transport of macromolecules. Here, we provide a composite multiscale structure of the yeast NPC, based on improved 3D density maps from cryogenic electron microscopy and AlphaFold2 models. Key features of the inner and outer rings were integrated into a comprehensive model. We resolved flexible connectors that tie together the core scaffold, along with equatorial transmembrane complexes and a lumenal ring that anchor this channel within the pore membrane. The organization of the nuclear double outer ring reveals an architecture that may be shared with ancestral NPCs. Additional connections between the core scaffold and the central transporter suggest that under certain conditions, a degree of local organization is present at the periphery of the transport machinery. These connectors may couple conformational changes in the scaffold to the central transporter to modulate transport. Collectively, this analysis provides insights into assembly, transport, and NPC evolution.
History
DepositionJul 20, 2023-
Header (metadata) releaseOct 11, 2023-
Map releaseOct 11, 2023-
UpdateOct 11, 2023-
Current statusOct 11, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_41300.png

Downloads & links

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Map

FileDownload / File: emd_41300.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotationc8 inner spoke ring recombined from multibody
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
2.66 Å/pix.
x 480 pix.
= 1276.8 Å		2.66 Å/pix.
x 480 pix.
= 1276.8 Å		2.66 Å/pix.
x 480 pix.
= 1276.8 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 2.66 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.65
Minimum - Maximum-1.8944535 - 5.8161745
Average (Standard dev.)0.01061699 (±0.09451712)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions480480480
Spacing480480480
CellA=B=C: 1276.8 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: spoke multibody half map

Fileemd_41300_additional_1.map
Annotationspoke multibody half map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: spoke multibody half map

Fileemd_41300_additional_2.map
Annotationspoke multibody half map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Additional map: c2 spoke map aligned within c8 ring and aligned with pdb

Fileemd_41300_additional_3.map
Annotationc2 spoke map aligned within c8 ring and aligned with pdb
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map of c8 spoke inner ring

Fileemd_41300_half_map_1.map
Annotationhalf map of c8 spoke inner ring
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map of c8 spoke inner ring

Fileemd_41300_half_map_2.map
Annotationhalf map of c8 spoke inner ring
Projections & Slices
AxesZYX

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Slices (1/2)
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Histogram (log scale)

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Sample components

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Entire : Nuclear Pore Complex

EntireName: Nuclear Pore Complex
Components
  • Complex: Nuclear Pore Complex
    • Protein or peptide: Spoke connector
    • Protein or peptide: Nucleoporin NUP170
    • Protein or peptide: Nucleoporin NUP157
    • Protein or peptide: Nucleoporin NSP1
    • Protein or peptide: Nucleoporin NUP57
    • Protein or peptide: Nucleoporin NUP49/NSP49
    • Protein or peptide: Nucleoporin NUP192
    • Protein or peptide: Nucleoporin NUP188
    • Protein or peptide: Nucleoporin NIC96
    • Protein or peptide: Nucleoporin NUP53
    • Protein or peptide: Nucleoporin 59

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Supramolecule #1: Nuclear Pore Complex

SupramoleculeName: Nuclear Pore Complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all / Details: Protein A tagged Mlp1 pullout of NPC
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Strain: MATa ade2-1 ura3-1 his3-11,15 trp1-1 leu2-3,112 can1-100 MLP1-PPX-ProteinA::HIS5
Organelle: nucleus / Location in cell: nuclear envelope
Molecular weightTheoretical: 18 MDa

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Macromolecule #1: Spoke connector

MacromoleculeName: Spoke connector / type: protein_or_peptide / ID: 1 / Number of copies: 18 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 3.166895 KDa
SequenceString:
(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)

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Macromolecule #2: Nucleoporin NUP170

MacromoleculeName: Nucleoporin NUP170 / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: MATa ade2-1 ura3-1 his3-11
Molecular weightTheoretical: 169.651969 KDa
SequenceString: MFQSFFHNNG PAAAGETFSD SRSYPLTNHQ EVPRNGLNEL ASSATKAQQQ PTHILNSYPI TGSNPLMRAS AMGATSGSIN PNMSNMNEH IRVSGMGTSK PLDLAGKYID HLQHKDSNTP VLDERSYYNS GVDYNFSREK NGLGAFTPFE KQDVFNIPDE I LHEFSTSQ ...String:
MFQSFFHNNG PAAAGETFSD SRSYPLTNHQ EVPRNGLNEL ASSATKAQQQ PTHILNSYPI TGSNPLMRAS AMGATSGSIN PNMSNMNEH IRVSGMGTSK PLDLAGKYID HLQHKDSNTP VLDERSYYNS GVDYNFSREK NGLGAFTPFE KQDVFNIPDE I LHEFSTSQ TKTDMGIFPE LNRCWITIDN KLILWNINND NEYQVVDDMK HTIQKVALVR PKPNTFVPAV KHLLLISTTM EL FMFAISL DKATNELSVF NTHLSVPVQG IDVIDIVSHE RSGRIFFAGQ ASGLNIWELH YSGSDDWFNS KCSKVCLTKS ALL SLLPTN MLSQIPGVDF IQALFEDNSN GNGGFSQETI TQLTIDQQRG IIYSLSSKST IRAYVITEKS LEGPMSIEPA YISR IIGTT TARAAPILGP KYLKIVKISS VAPEENNNLF LVALTVGGVR LYFNGSMGRF NIEALRLESI KFPPSSVTPE VIQQE LLHQ QQEQAKRSFP FFSNLMSSEP VLLKFQKKSS VLLETTKAST IISPGIFFSA VIKSSQQTHQ QEKKENSSVT GTTATA GSK TVKQQPVTLQ HKLFVSVPDY GILKTHGKYV ENATFLETAG PVQQIIPLSG LFNATTKPQG FANEFATQYT SETLRVA VL TSTSIEIYKY RTPDEIFEDL IDNPLPFVLN YGAAEACSTA LFVTCKSNKS EKLRSNALTF LTMGIPGVVD IKPVYNRY S VSTVSSLLSK PTLSTATTNL QQSITGFSKP SPANKEDFDL DDVILSPRFY GIALLITRLL RDIWGRHVFM TFTDNRVTS HAFISSSDPI TPSINNLKSD EISQNRNIIS KVSISKDCIE YYLSSINILN EFFITYGDSI SQISAPYVLA NNSNGRVIDK TEEVANQAE SIAINAMIKM VQSIKEGLSF LNVLYEESEV EGFDNQYLGF KDIISFVSLD VQKDLVKLDF KDLFAPNDKT K SLIREILL SIINRNITKG ASIEYTATAL QERCGSFCSA SDILGFRAIE HLRRAKEIGL RNYDSLNYHL KNATALLEQI VD DLSIEKL KEAVSMMLSV NYYPKSIEFL LNIANSMDKG KLACQYVANG FLENDDRKQY YDKRILVYDL VFDTLIKVDE LAE KKQSSK TQNQISISND DEVKLRQKSY EAALKYNDRL FHYHMYDWLV SQNREEKLLD IETPFILPYL MEKAGSSLKI SNIL WVYYS RRSKFFESAE ILYRLATSNF DITLFERIEF LSRANGFCNS VSPLSQKQRI VQLASRIQDA CEVAGIQGDI LSLVY TDAR IDSAIKDELI KTLDGKILST SELFNDFAVP LSYHEIALFI FKIADFRDHE VIMAKWDELF QSLRMEFNNT GKKEDS MNF INLLSNVLIK IGKNVQDSEF IFPIFELFPI VCNFFYETLP KEHIVSGSIV SIFITAGVSF NKMYYILKEL IETSDSD NS VFNKEMTWLI HEWYKSDRKF RDIISYNDII HLKEYKIDND PIEKYVKNSG NNLGICFYKE

UniProtKB: Nucleoporin NUP170

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Macromolecule #3: Nucleoporin NUP157

MacromoleculeName: Nucleoporin NUP157 / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: 15 trp1-1 leu2-3
Molecular weightTheoretical: 156.827484 KDa
SequenceString: MYSTPLKKRI DYDRETFTAS ASLGGNRLRN RPRDDQNNGK PNLSSRSFLS ERKTRKDVLN KYGEAGNTIE SELRDVTTHV KISGLTSSE PLQLASEFVQ DLSFRDRNTP ILDNPDYYSK GLDYNFSDEV GGLGAFTPFQ RQQVTNIPDE VLSQVSNTEI K SDMGIFLE ...String:
MYSTPLKKRI DYDRETFTAS ASLGGNRLRN RPRDDQNNGK PNLSSRSFLS ERKTRKDVLN KYGEAGNTIE SELRDVTTHV KISGLTSSE PLQLASEFVQ DLSFRDRNTP ILDNPDYYSK GLDYNFSDEV GGLGAFTPFQ RQQVTNIPDE VLSQVSNTEI K SDMGIFLE LNYCWITSDN KLILWNINNS SEYHCIDEIE HTILKVKLVK PSPNTFVSSV ENLLIVATLF DIYILTISFN DR THELNIF NTGLKVNVTG FNVSNIISYE RTGQIFFTGA TDGVNVWELQ YNCSENLFNS KSNKICLTKS NLANLLPTKL IPS IPGGKL IQKVLEGDAG TEEETISQLE VDQSRGVLHT LSTKSIVRSY LITSNGLVGP VLIDAAHIRR GMNALGVKNS PLLS NRAFK IAKIVSISMC ENNDLFLAVI TTTGVRLYFK GSISRRSIGS LKLDSVKFPP TSISSSLEQN KSFIIGHHPL NTHDT GPLS TQKASSTYIN TTCASTIISP GIYFTCVRKR ANSGELSKGI TNKALLENKE EHKLYVSAPD YGILKNYGKY VENTAL LDT TDEIKEIVPL TRSFNYTSTP QGYANVFASQ YSAEPLKVAV LTSNALEIYC YRTPDEVFES LIENPLPFIH SYGLSEA CS TALYLACKFN KSEHIKSSAL AFFSAGIPGV VEIKPKSSRE SGSVPPISQN LFDKSGECDG IVLSPRFYGS ALLITRLF S QIWEERVFVF KRASKTEKMD AFGISITRPQ VEYYLSSISV LADFFNIHRP SFVSFVPPKG SNAITASDAE SIAMNALIL LINSIKDALS LINVFYEDID AFKSLLNTLM GAGGVYDSKT REYFFDLKFH DLFTPNAKTK QLIKEILIEV VNANIASGTS ADYIVNVLK ERFGSFCHSA DILCYRAGEH LEAAQKFEMI DSKISRNHLD TAIDLYERCA ENIELCELRR VVDIMVKLNY Q PKTVGFLL RFADKIDKGN QAQEYVSRGC NTADPRKVFY DKRINVYTLI FEIVKSVDDY TSIEQSPSIA NISIFSPASS LK KRVYSVI MNSNNRFFHY CFYDWLVANK RQDYLLRLDS QFVLPYLKER AEKSLEISNL LWFYLFKEEH FLEAADVLYA LAS SDFDLK LSERIECLAR ANGLCDSSTS FDQKPALVQL SENIHELFDI ASIQDDLLNL VRNETRIDED YRKQLTLKLN GRVL PLSDL FNDCADPLDY YEIKLRIFKV SQFKDEKVIQ GEWNRLLDSM KNAPSPDVGS VGQESFLSSI SNTLIRIGKT TRDTD VVFP VHFLMNKILE SFIDKSSAAD GSVCSMFLLA GVSHLKLYYI LSRIIENSEG NVELAKKEMV WLIKDWYQSD SDLRGS IAP EQIKKLEKYD PNTDPVQDYV KDRHHGLK

UniProtKB: Nucleoporin NUP157

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Macromolecule #4: Nucleoporin NSP1

MacromoleculeName: Nucleoporin NSP1 / type: protein_or_peptide / ID: 4 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: 112 can1-100 MLP1-PPX-ProteinA::HIS5
Molecular weightTheoretical: 86.611672 KDa
SequenceString: MNFNTPQQNK TPFSFGTANN NSNTTNQNSS TGAGAFGTGQ STFGFNNSAP NNTNNANSSI TPAFGSNNTG NTAFGNSNPT SNVFGSNNS TTNTFGSNSA GTSLFGSSSA QQTKSNGTAG GNTFGSSSLF NNSTNSNTTK PAFGGLNFGG GNNTTPSSTG N ANTSNNLF ...String:
MNFNTPQQNK TPFSFGTANN NSNTTNQNSS TGAGAFGTGQ STFGFNNSAP NNTNNANSSI TPAFGSNNTG NTAFGNSNPT SNVFGSNNS TTNTFGSNSA GTSLFGSSSA QQTKSNGTAG GNTFGSSSLF NNSTNSNTTK PAFGGLNFGG GNNTTPSSTG N ANTSNNLF GATANANKPA FSFGATTNDD KKTEPDKPAF SFNSSVGNKT DAQAPTTGFS FGSQLGGNKT VNEAAKPSLS FG SGSAGAN PAGASQPEPT TNEPAKPALS FGTATSDNKT TNTTPSFSFG AKSDENKAGA TSKPAFSFGA KPEEKKDDNS SKP AFSFGA KSNEDKQDGT AKPAFSFGAK PAEKNNNETS KPAFSFGAKS DEKKDGDASK PAFSFGAKPD ENKASATSKP AFSF GAKPE EKKDDNSSKP AFSFGAKSNE DKQDGTAKPA FSFGAKPAEK NNNETSKPAF SFGAKSDEKK DGDASKPAFS FGAKS DEKK DSDSSKPAFS FGTKSNEKKD SGSSKPAFSF GAKPDEKKND EVSKPAFSFG AKANEKKESD ESKSAFSFGS KPTGKE EGD GAKAAISFGA KPEEQKSSDT SKPAFTFGAQ KDNEKKTEES STGKSTADVK SSDSLKLNSK PVELKPVSLD NKTLDDL VT KWTNQLTESA SHFEQYTKKI NSWDQVLVKG GEQISQLYSD AVMAEHSQNK IDQSLQYIER QQDELENFLD NFETKTEA L LSDVVSTSSG AAANNNDQKR QQAYKTAQTL DENLNSLSSN LSSLIVEINN VSNTFNKTTN IDINNEDENI QLIKILNSH FDALRSLDDN STSLEKQINS IKK

UniProtKB: Nucleoporin NSP1

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Macromolecule #5: Nucleoporin NUP57

MacromoleculeName: Nucleoporin NUP57 / type: protein_or_peptide / ID: 5 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 57.547145 KDa
SequenceString: MFGFSGSNNG FGNKPAGSTG FSFGQNNNNT NTQPSASGFG FGGSQPNSGT ATTGGFGANQ ATNTFGSNQQ SSTGGGLFGN KPALGSLGS SSTTASGTTA TGTGLFGQQT AQPQQSTIGG GLFGNKPTTT TGGLFGNSAQ NNSTTSGGLF GNKVGSTGSL M GGNSTQNT ...String:
MFGFSGSNNG FGNKPAGSTG FSFGQNNNNT NTQPSASGFG FGGSQPNSGT ATTGGFGANQ ATNTFGSNQQ SSTGGGLFGN KPALGSLGS SSTTASGTTA TGTGLFGQQT AQPQQSTIGG GLFGNKPTTT TGGLFGNSAQ NNSTTSGGLF GNKVGSTGSL M GGNSTQNT SNMNAGGLFG AKPQNTTATT GGLFGSKPQG STTNGGLFGS GTQNNNTLGG GGLFGQSQQP QTNTAPGLGN TV STQPSFA WSKPSTGSNL QQQQQQQIQV PLQQTQAIAQ QQQLSNYPQQ IQEQVLKCKE SWDPNTTKTK LRAFVYNKVN ETE AILYTK PGHVLQEEWD QAMEKKPSPQ TIPIQIYGFE GLNQRNQVQT ENVAQARIIL NHILEKSTQL QQKHELDTAS RILK AQSRN VEIEKRILKL GTQLATLKNR GLPLGIAEEK MWSQFQTLLQ RSEDPAGLGK TNELWARLAI LKERAKNISS QLDSK LMVF NDDTKNQDSM SKGTGEESND RINKIVEILT NQQRGITYLN EVLEKDAAIV KKYKNKT

UniProtKB: Nucleoporin NUP57

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Macromolecule #6: Nucleoporin NUP49/NSP49

MacromoleculeName: Nucleoporin NUP49/NSP49 / type: protein_or_peptide / ID: 6 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 49.174762 KDa
SequenceString: MFGLNKASST PAGGLFGQAS GASTGNANTG FSFGGTQTGQ NTGPSTGGLF GAKPAGSTGG LGASFGQQQQ QSQTNAFGGS ATTGGGLFG NKPNNTANTG GGLFGANSNS NSGSLFGSNN AQTSRGLFGN NNTNNINNSS SGMNNASAGL FGSKPAGGTS L FGNTSTSS ...String:
MFGLNKASST PAGGLFGQAS GASTGNANTG FSFGGTQTGQ NTGPSTGGLF GAKPAGSTGG LGASFGQQQQ QSQTNAFGGS ATTGGGLFG NKPNNTANTG GGLFGANSNS NSGSLFGSNN AQTSRGLFGN NNTNNINNSS SGMNNASAGL FGSKPAGGTS L FGNTSTSS APAQNQGMFG AKPAGTSLFG NNAGNTTTGG GLFGSKPTGA TSLFGSSNNN NNNNNSNNIM SASGGLFGNQ QQ QLQQQPQ MQCALQNLSQ LPITPMTRIS ELPPQIRQEI EQLDQYIQKQ VQISHHLKAD TIDHDELIDS IPRDVAYLLK SES ATSQYL KQDLKKISSF KSLIDEDLLD TQTFSVLLQQ LLTPGSKISS NDLDKFFQKK IHLYEKKLED YCRILSDIET AVNG IDTDL FGAPNNPNST AITADLGSSE AENLLQLKTG LAAIVSTVIE EFTLFMDIAE RIAVLHQKTK TLASLSI

UniProtKB: Nucleoporin NUP49/NSP49

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Macromolecule #7: Nucleoporin NUP192

MacromoleculeName: Nucleoporin NUP192 / type: protein_or_peptide / ID: 7 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 191.718125 KDa
SequenceString: MKWSAIPFQT LYRSIESGEF DFDLFKEVLP DLQNLNLNTD KLKNNASRSQ LEKGEIELSD GSTFKVNQEF IFEAISLSDE LNLDEIVAC ELILSGDTTA NNGKVQYFLR RQYILQIVSF IVNCFHEDTE LYQELIKNGA LVSNILSAFK FIHTQLSEIK Q QINKAQIL ...String:
MKWSAIPFQT LYRSIESGEF DFDLFKEVLP DLQNLNLNTD KLKNNASRSQ LEKGEIELSD GSTFKVNQEF IFEAISLSDE LNLDEIVAC ELILSGDTTA NNGKVQYFLR RQYILQIVSF IVNCFHEDTE LYQELIKNGA LVSNILSAFK FIHTQLSEIK Q QINKAQIL ENYNALFQQN IKFRRDFLLR EYDILSQILY GLVDKGAIMK NKDFILSLLH HVSELDSNDF FIIYYTPAFF HL FASLRVL PDADVKLLHS QFMKDLKDDS IYTKPVKVAL IFIFFAYFIG WCKEDPKRRA DTMDFKTDVD EPMTSAVELG AIE QILIFA ADTSIVEQDK SMELFYDIRS LLERHIPRLI PKQLLDDEKI FSQTTNSTYN PASATDNMSG RGLWNPSYPG MMST TGTAR LNSMPNNVNE YSYTTIVLSD QTQEFFLSSF DDVLQTIITD CAFLLTKIKD AEEDSLLSGE DLTLDDISLK ADLER FFLS IYFFYASRPE YSCTFWSDKE SNAYGFIEWC SRCNDNLMRS CFYLMVSSLS FGPENALNVY HYFGENSSIS WKNIAQ CLS DYTKKISNFN SSLHKRQQFS ESTHNDIDST AVALEEGLNE EAVIFLSSLL TLVGSVTYQV DEDVKSSLSK VFSDVLF EF TKINTPLVGA AFKVISNLVP KLESSRTKFW SFLDSLIFKD SSLNYSSESY RNAFTNVLTK YSDVLGFLQL FHNLISIH S RENNSEYMVF GKLAFPTRLG QGYRKVGIWP YFDYIFNDIL AHVDQIVDIR NKRAVQLPIL KIIYTGLCSF DYSVILNSI PAAANLDALV DCENFFNYVQ ECPAIPIFNY IFTEKIYKSI FNVVDVGVDQ LSIELEGGKN QAELLQLAVK IINKVLDYQE TYVEELFPI VKKHGKTDYF LPKNYSLHGL RSFYDAIFFN IPLVAHLGLY VGVDDQILAT NSLRILAKLS ERSNGSVASL S KRNKLLTI FDSVDESARI KDAFITQLES SITDAGVLAL KLELLDFLTS NLSNYSRTMT ISHLLLGFQV SNVISLGPNL AT FISSGTS LLDSLISVLE ASLNSITKDN IDYAPMRLAT AALEIILKLC RNPLTSGLLY SYLIKENFFE RIMILDPQVT RFT TWNGSP FDNSTEEKCK NFIESESVGA FLSFLAYRNY WTQYLGLFIH KISFSGTKSE VLTYVNYLIS NTMYSVRLFS FLDP LNYGN ICEPKETLSI FTNVPLNLEQ VTLNKYCSGN IYDFHKMENL MRLIKRVRAE SLHSNSFSLT VSKEQFLKDA DVECI KAKS HFTNIISRNK ALELNLSVLH SWVQLVQIIV TDGKLEPSTR SNFILEVFGT IIPKISDYIE FNITFSEELV SLAVFL FDI YNRDRKLITD KGTVDGRLYQ LFKTCIQGIN SPLSSVALRS DFYILANHYL SRVLSDQVGS EKVLQDLRLG SKKLVEI IW NDVVYGEGTS RVTGILLLDS LIQLANRSKE NFILDSLMKT TRLLLIIRSL KNTDALLNST TEHINIDDLL YELTAFKA T VFFLIRVAET RGGASALIEN NLFRIIAELS FLKVDPDLGL DLMFDEVYVQ NSKFLKVNVT LDNPLLVDKD ANGVSLFEL IVPIFQLISA VLVSMGSSNK AVVQTVKGLL NTYKRLVIGI FKRDLLREKE DKKNSSDPNN QSLNEMVKLI VMLCTLTGYQ NND

UniProtKB: Nucleoporin NUP192

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Macromolecule #8: Nucleoporin NUP188

MacromoleculeName: Nucleoporin NUP188 / type: protein_or_peptide / ID: 8 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 188.753281 KDa
SequenceString: MATPSFGNSS PQLTFTHVAN FMNDAAADVS AVDAKQLAQI RQFLKANKTN LIESLNTIRQ NVTSSGDHNK LRSTIANLLQ INVDNDPFF AQSEDLSHAV EFFMSERSSR LHIVYSLLVN PDIDLETYSF IDNDRFNVVG KLISIISSVI QNYDIITASS L AHDYNNDQ ...String:
MATPSFGNSS PQLTFTHVAN FMNDAAADVS AVDAKQLAQI RQFLKANKTN LIESLNTIRQ NVTSSGDHNK LRSTIANLLQ INVDNDPFF AQSEDLSHAV EFFMSERSSR LHIVYSLLVN PDIDLETYSF IDNDRFNVVG KLISIISSVI QNYDIITASS L AHDYNNDQ DMFTIVSLVQ LKKFSDLKFI LQILQILNLM ILNTKVPVDI VNQWFLQYQN QFVEFCRNIN STDKSIDTSS LQ LYKFQNF QDLSYLSETL ISRISSLFTI TTILILGLNT SIAQFDIQSP LYMDTETFDT VNSALENDVA TNIVNEDPIF HPM IHYSWS FILYYRRALQ SSESFDDSDI TKFALFAESH DVLQKLNTLS EILSFDPVYT TVITVFLEFS LNFIPITAST SRVF AKIIS KAPEQFIENF LTNDTFEKKL SIIKAKLPLL NESLIPLINL ALIDTEFANF ELKDICSFAV TKSSLNDLDY DLIAD TITN SSSSSDIIVP DLIELKSDLL VAPPLENENS NCLLSIPKST KGKILTIKQQ QQQQQQQNGQ QPPTTSNLII FLYKFN GWS LVGRILQNLL HSYMEKGTQL DDLQHELMIS IIKLVTNVVD PKTSIEKSSE ILSYLSNSLD TSASTINGAS IIQVIFE IF EISLQRKDYT SIVQCCEFMT MLTPNYLHLV SSYLNKSDLL DKYGKTGLSN MILGSVELST GDYTFTIQLL KLTKVFIR E SLSLKNIHIS KRSKIDIINK LILHAIHIFE SYYNWKYNNF LQKFEIAFHL TLIFYDVLHD VFTINPHQKD QLIISSSAN KLLQLFLTPM DSIDLAPNTL TNILISPLNT TTKILGDKIL GNLYSKVMNN SFKLCTLLIA IRGSNRDLKP SNLEKLLFIN SSKLVDVYT LPSYVHFKVQ IIELLSYLVE APWNDDYPFL LSFLGEAKSM AFLKEVLSDL SSPVQDWNLL RSLYIFFTTL L ESKQDGLS ILFLTGQFAS NKKINDESSI DKKSSILTVL QKNSLLLDST PEEVSCKLLE TITYVLNTWT NSKIFIKDPK FV NSLLAKL KDSKKLFQKK ENLTRDETVS LIKKYKLISR IVEIFALCIY NSTDSNSEIL NFLNQEDLFE LVHHFFQIDG FNK TFHDEL NLKFKEKWPS LELQSFQKIP LSRINENENF GYDIPLLDIV LKADRSWNEP SKSQTNFKEE ITDASLNLQY VNYE ISTAK AWGALITTFV KRSTVPLNDG FVDLVEHFLK LNIDFGSDKQ MFTQIYLERI ELSFYILYSF KLSGKLLKEE KIIEL MNKI FTIFKSGEID FIKNIGKSLK NNFYRPLLRS VLVLLELVSS GDRFIELISD QLLEFFELVF SKGVYLILSE ILCQIN KCS TRGLSTDHTT QIVNLEDNTQ DLLLLLSLFK KITNVNPSKN FNVILASSLN EVGTLKVILN LYSSAHLIRI NDEPILG QI TLTFISELCS IEPIAAKLIN SGLYSVLLES PLSVAIQQGD IKPEFSPRLH NIWSNGLLSI VLLLLSQFGI KVLPETCL F VSYFGKQIKS TIYNWGDNKL AVSSSLIKET NQLVLLQKML NLLNYQELFI QPKNSDDQQE AVELVIGLDS EHDKKRLSA ALSKFLTHPK YLNSRIIPTT LEEQQQLEDE SSRLEFVKGI SRDIKALQDS LFKDV

UniProtKB: Nucleoporin NUP188

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Macromolecule #9: Nucleoporin NIC96

MacromoleculeName: Nucleoporin NIC96 / type: protein_or_peptide / ID: 9 / Number of copies: 4 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 96.291586 KDa
SequenceString: MLETLRGNKL HSGTSKGANK KLNELLESSD NLPSASSELG SIQVSINELR RRVFQLRSKN KASKDYTKAH YLLANSGLSF EDVDAFIKD LQTNQFLEPN PPKIIESEEL EFYIRTKKEE NILMSIEQLL NGATKDFDNF INHNLNLDWA QHKNEVMKNF G ILIQDKKT ...String:
MLETLRGNKL HSGTSKGANK KLNELLESSD NLPSASSELG SIQVSINELR RRVFQLRSKN KASKDYTKAH YLLANSGLSF EDVDAFIKD LQTNQFLEPN PPKIIESEEL EFYIRTKKEE NILMSIEQLL NGATKDFDNF INHNLNLDWA QHKNEVMKNF G ILIQDKKT VDHKKSISSL DPKLPSWGNK GNNILNSNES RLNVNENNIL REKFENYARI VFQFNNSRQA NGNFDIANEF IS ILSSANG TRNAQLLESW KILESMKSKD INIVEVGKQY LEQQFLQYTD NLYKKNMNEG LATNVNKIKS FIDTKLKKAD KSW KISNLT VINGVPIWAL IFYLLRAGLI KEALQVLVEN KANIKKVEQS FLTYFKAYAS SKDHGLPVEY STKLHTEYNQ HIKS SLDGD PYRLAVYKLI GRCDLSRKNI PAVTLSIEDW LWMHLMLIKE KDAENDPVYE RYSLEDFQNI IISYGPSRFS NYYLQ TLLL SGLYGLAIDY TYTFSEMDAV HLAIGLASLK LFKIDSSTRL TKKPKRDIRF ANILANYTKS FRYSDPRVAV EYLVLI TLN EGPTDVELCH EALRELVLET KEFTVLLGKI GRDGARIPGV IEERQPLLHV RDEKEFLHTI TEQAARRADE DGRIYDS IL LYQLAEEYDI VITLVNSLLS DTLSASDLDQ PLVGPDDNSE TNPVLLARRM ASIYFDNAGI SRQIHVKNKE ICMLLLNI S SIRELYFNKQ WQETLSQMEL LDLLPFSDEL SARKKAQDFS NLDDNIVKNI PNLLIITLSC ISNMIHILNE SKYQSSTKG QQIDSLKNVA RQCMIYAGMI QYRMPRETYS TLINIDVSL

UniProtKB: Nucleoporin NIC96

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Macromolecule #10: Nucleoporin NUP53

MacromoleculeName: Nucleoporin NUP53 / type: protein_or_peptide / ID: 10 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 52.688668 KDa
SequenceString: MADLQKQENS SRFTNVSVIA PESQGQHEQQ KQQEQLEQQK QPTGLLKGLN GFPSAPQPLF MEDPPSTVSG ELNDNPAWFN NPRKRAIPN SIIKRSNGQS LSPVRSDSAD VPAFSNSNGF NNVTFGSKKD PRILKNVSPN DNNSANNNAH SSDLGTVVFD S NEAPPKTS ...String:
MADLQKQENS SRFTNVSVIA PESQGQHEQQ KQQEQLEQQK QPTGLLKGLN GFPSAPQPLF MEDPPSTVSG ELNDNPAWFN NPRKRAIPN SIIKRSNGQS LSPVRSDSAD VPAFSNSNGF NNVTFGSKKD PRILKNVSPN DNNSANNNAH SSDLGTVVFD S NEAPPKTS LADWQKEDGI FSSKTDNIED PNLSSNITFD GKPTATPSPF RPLEKTSRIL NFFDKNTKTT PNTASSEASA GS KEGASTN WDDHAIIIFG YPETIANSII LHFANFGEIL EDFRVIKDFK KLNSKNMSKS PSLTAQKYPI YTGDGWVKLT YKS ELSKSR ALQENGIIMN GTLIGCVSYS PAALKQLASL KKSEEIINNK TSSQTSLSSK DLSNYRKTEG IFEKAKAKAV TSKV RNAEF KVSKNSTSFK NPRRLEIKDG RSLFLRNRGK IHSGVLSSIE SDLKKREQAS KSKKSWLNRL NNWLFGWNDL

UniProtKB: Nucleoporin NUP53

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Macromolecule #11: Nucleoporin 59

MacromoleculeName: Nucleoporin 59 / type: protein_or_peptide / ID: 11 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 58.853902 KDa
SequenceString: MFGIRSGNNN GGFTNLTSQA PQTTQMFQSQ SQLQPQPQPQ PQQQQQHLQF NGSSDASSLR FGNSLSNTVN ANNYSSNIGN NSINNNNIK NGTNNISQHG QGNNPSWVNN PKKRFTPHTV IRRKTTKQNS SSDINQNDDS SSMNATMRNF SKQNQDSKHN E RNKSAANN ...String:
MFGIRSGNNN GGFTNLTSQA PQTTQMFQSQ SQLQPQPQPQ PQQQQQHLQF NGSSDASSLR FGNSLSNTVN ANNYSSNIGN NSINNNNIK NGTNNISQHG QGNNPSWVNN PKKRFTPHTV IRRKTTKQNS SSDINQNDDS SSMNATMRNF SKQNQDSKHN E RNKSAANN DINSLLSNFN DIPPSVTLQD WQREDEFGSI PSLTTQFVTD KYTAKKTNRS AYDSKNTPNV FDKDSYVRIA NI EQNHLDN NYNTAETNNK VHETSSKSSS LSAIIVFGYP ESISNELIEH FSHFGHIMED FQVLRLGRGI NPNTFRIFHN HDT GCDEND STVNKSITLK GRNNESNNKK YPIFTGESWV KLTYNSPSSA LRALQENGTI FRGSLIGCIP YSKNAVEQLA GCKI DNVDD IGEFNVSMYQ NSSTSSTSNT PSPPNVIITD GTLLREDDNT PAGHAGNPTN ISSPIVANSP NKRLDVIDGK LPFMQ NAGP NSNIPNLLRN LESKMRQQEA KYRNNEPAGF THKLSNWLFG WNDL

UniProtKB: Nucleoporin ASM4

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration0.1 mg/mL
BufferpH: 7.5
Details: 20mM HEPES,50mM Potassium acetate,20mM NaCl,2mM MgCl2,1mM DTT
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 283 K / Instrument: FEI VITROBOT MARK III
DetailsOne step affinity purified

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsEnergy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
DetailsPreliminary grid screening done manually with individual images of low magnification montages of candidate meshes.
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 2-40 / Number grids imaged: 1 / Number real images: 4015 / Average electron dose: 40.0 e/Å2 / Details: 3218 images retained after triage
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Calibrated magnification: 37651 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.5 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 26049
Startup modelType of model: OTHER
Details: C8 symmetrized NPC map segmented in CHIMERA with Segger to provide initial 3D map of the double outer ring, which was low pass filtered for the first reference.
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 6.6 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 3.3.2)
Details: Multibody extracted spoke image stack processed in CryoSPARC
Number images used: 208392
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0.7) / Details: C1 symmetry used with Multibody particles
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC (ver. 3.3.2) / Software - details: non-uniform refinement
Details: C1 symmetry used to refine inner ring spoke with appropriate mask
Final 3D classificationSoftware - Name: RELION (ver. 3.0.7)
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelChain - Source name: Other / Chain - Initial model type: experimental model / Details: none
DetailsRigid body docking of 3D map of Nucleoporins in the spoke complex with Chimera followed by flexible fitting and rebuilding in Coot.
RefinementSpace: REAL / Protocol: FLEXIBLE FIT
Output model

PDB-8tj5:
Inner spoke ring of the yeast NPC

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