2020-07-15 REL PDBe PDBe PDBe 2020-04-06 2020-05-20 2020-05-20 2020-07-15 Cancer Research UK C47547/A21536 United Kingdom Wellcome Trust 200818/Z/16/Z United Kingdom Cutts EE Beuron F Morris E Vannini A Kong M Cutts EE Pan D Beuron F Kaliyappan T Xue C Morris EP Musacchio A Vannini A Greene EC Mol.Cell US 79 99 114.e9 2020 32445620 doi:10.1016/j.molcel.2020.04.026 1097-2765 2168 MOCEFL EMD-10827 associated EM volume Condensin I pentameric complex of SMC2, SMC4, CapH, CapD3 and CapG Condensin I pentameric complex of SMC2, SMC4, CapH, CapD3 and CapG 0 1 2 3 4 5 Homo sapiens Trichoplusia ni 0.643 SMC2 Homo sapiens Trichoplusia ni LEVO HIKSIILEGFKSYAQRTEV NGFDPLFNAITGLNGSGKSN ILDSICFLLGISNLSQVRAS NLQDLVYKNGQAGITKASVS ITFDNSDKKQSPLGFEVHDE ITVTRQVVIGGRNKYLINGV NANNTRVQDLFCSVGLNVNN PHFLIMQGRITKVLNMKPPE ILSMIEEAAGTRMYEYKKIA AQKTIEKKEAKLKEIKTILE EEITPTIQKLKEERSSYLEY QKVMREIEHLSRLYIAYQFL LAEDTKVRSAEELKEMQDKV IKLQEELSENDKKIKALNHE IEELEKRKDKETGGILRSLE DALAEAQRVNTKSQSAFDLK KKNLACEESKRKELEKNMVE DSKTLAAKEKEVKKITDGLH ALQEASNKDAEALAAAQQHF NAVSAGLSSNEDGAEATLAG QMMACKNDISKAQTEAKQAQ MKLKHAQQELKNKQAEVKKM DSGYRKDQEALEAVKRLKEK LEAEMKKLNYEENKEESLLE KRRQLSRDIGRLKETYEALL ARFPNLRFAYKDPEKNWNRN CVKGLVASLISVKDTSATTA LELVAGERLYNVVVDTEVTG KKLLERGELKRRYTIIPLNK ISARCIAPETLRVAQNLVGP DNVHVALSLVEYKPELQKAM EFVFGTTFVCDNMDNAKKVA FDKRIMTRTVTLGGDVFDPH GTLSGGARSQAASILTKFQE LKDVQDELRIKENELRALEE ELAGLKNTAEKYRQLKQQWE MKTEEADLLQTKLQQSSYHK QQEELDALKKTIEESEETLK NTKEIQRKAEEKYEVLENKM KNAEAERERELKDAQKKLDC AKTKADASSKKMKEKQQEVE AITLELEELKREHTSYKQQL EAVNEAIKSYESQIEVMAAE VAKNKESVNKAQEEVTKQKE VITAQDTVIKAKYAEVAKHK EQNNDSQLKIKELDHNISKH KREAEDGAAKVSKMLKDYDW INAERHLFGQPNSAYDFKTN NPKEAGQRLQKLQEMKEKLG RNVNMRAMNVLTEAEERYND LMKKKRIVENDKSKILTTIE DLDQKKNQALNIAWQKVNKD FGSIFSTLLPGANAMLAPPE GQTVLDGLEFKVALGNTWKE NLTELSGGQRSLVALSLILS MLLFKPAPIYILDEVDAALD LSHTQNIGQMLRTHFTHSQF IVVSLKEGMFNNANVLFKTK FVDGVSTVARFTQCQNGKIS KEAKSKAKPPKGAHVEV SMC4 Homo sapiens Trichoplusia ni LEVO MAHHHHHHHHHHLEVLFQGP RKGTQPSTARRREEGPPPPS PDGASSDAEPEPPSGRTESP ATAAETASEELDNRSLEEIL NSIPPPPPPAMTNEAGAPRL MITHIVNQNFKSYAGEKILG PFHKRFSCIIGPNGSGKSNV IDSMLFVFGYRAQKIRSKKL SVLIHNSDEHKDIQSCTVEV HFQKIIDKEGDDYEVIPNSN FYVSRTACRDNTSVYHISGK KKTFKDVGNLLRSHGIDLDH NRFLILQGEVEQIAMMKPKG QTEHDEGMLEYLEDIIGCGR LNEPIKVLCRRVEILNEHRG EKLNRVKMVEKEKDALEGEK NIAIEFLTLENEIFRKKNHV CQYYIYELQKRIAEMETQKE KIHEDTKEINEKSNILSNEM KAKNKDVKDTEKKLNKITKF IEENKEKFTQLDLEDVQVRE KLKHATSKAKKLEKQLQKDK EKVEEFKSIPAKSNNIINET TTRNNALEKEKEKEEKKLKE VMDSLKQETQGLQKEKESRE KELMGFSKSVNEARSKMDVA QSELDIYLSRHNTAVSQLTK AKEALIAASETLKERKAAIR DIEGKLPQTEQELKEKEKEL QKLTQEETNFKSLVHDLFQK VEEAKSSLAMNRSRGKVLDA IIQEKKSGRIPGIYGRLGDL GAIDEKYDVAISSCCHALDY IVVDSIDIAQECVNFLKRQN IGVATFIGLDKMAVWAKKMT EIQTPENTPRLFDLVKVKDE KIRQAFYFALRDTLVADNLD QATRVAYQKDRRWRVVTLQG QIIEQSGTMTGGGSKVMKGR MGSSLVIEISEEEVNKMESQ LQNDSKKAMQIQEQKVQLEE RVVKLRHSEREMRNTLEKFT ASIQRLIEQEEYLNVQVKEL EANVLATAPDKKKQKLLEEN VSAFKTEYDAVAEKAGKVEA EVKRLHNTIVEINNHKLKAQ QDKLDKINKQLDECASAITK AQVAIKTADRNLQKAQDSVL RTEKEIKDTEKEVDDLTAEL KSLEDKAAEVVKNTNAAEES LPEIQKEHRNLLQELKVIQE NEHALQKDALSIKLKLEQID GHIAEHNSKIKYWHKEISKI SLHPIEDNPIEEISVLSPED LEAIKNPDSITNQIALLEAR CHEMKPNLGAIAEYKKKEEL YLQRVAELDKITYERDSFRQ AYEDLRKQRLNEFMAGFYII TNKLKENYQMLTLGGDAELE LVDSLDPFSEGIMFSVRPPK KSWKKIFNLSGGEKTLSSLA LVFALHHYKPTPLYFMDEID AALDFKNVSIVAFYIYEQTK NAQFIIISLRNNMFEISDRL IGIYKTYNITKSVAVNPKEI ASKGLC CapH Homo sapiens Trichoplusia ni LEVO MGPPGPALPATMNNSSSETR GHPHSASSPSERVFPMPLPR KAPLNIPGTPVLEDFPQNDD EKERLQRRRSRVFDLQFSTD SPRLLASPSSRSIDISATIP KFTNTQITEHYSTCIKLSTE NKITTKNAFGLHLIDFMSEI LKQKDTEPTNFKVAAGTLDA STKIYAVRVDAVHADVYRVL GGLGKDAPSLEEVEGHVADG SATEMGTTKKAVKPKKKHLH RTIEQNINNLNVSEADRKCE IDPMFQKTAASFDECSTAGV FLSTLHCQDYRSELLFPSDV QTLSTGEPLELPELGCVEMT DLKAPLQQCAEDRQICPSLA GFQFTQWDSETHNESVSALV DKFKKNDQVFDINAEVDESD CGDFPDGSLGDDFDANDEPD HTAVGDHEEFRSWKEPCQVQ SCQEEMISLGDGDIRTMCPL LSMKPGEYSYFSPRTMSMWA GPDHWRFRPRRKQDAPSQSE NKKKSTKKDFEIDFEDDIDF DVYFRKTKAATILTKSTLEN QNWRATTLPTDFNYNVDTLV QLHLKPGTRLLKMAQGHRVE TEHYEEIEDYDYNNPNDTSN FCPGLQAADSDDEDLDDLFV GPVGNSDLSPYPCHPPKTAQ QNGDTPEAQGLDITTYGESN LVAEPQKVNKIEIHYAKTAK KMDMKKLKQSMWSLLTALSG KEADAEANHREAGKEAALAE VADEKMLSGLTKDLQRSLPP VMAQNLSIPLAFACLLHLAN EKNLKLEGTEDLSDVLVRQG DENLYFQSWSHPQFEKGGGS GGGSGGGSWSHPQFEK CapD2 Homo sapiens Trichoplusia ni LEVO MAPQMYEFHLPLSPEELLKS GGVNQYVVQEVLSIKHLPPQ LRAFQAAFRAQGPLAMLQHF DTIYSILHHFRSIDPGLKED TLQFLIKVVSRHSQELPAIL DDTTLSGSDRNAHLNALKMN CYALIRLLESFETMASQTNL VDLDLGGKGKKARTKAAHGF DWEEERQPILQLLTQLLQLD IRHLWNHSIIEEEFVSLVTG CCYRLLENPTINHQKNRPTR EAITHLLGVALTRYNHMLSA TVKIIQMLQHFEHLAPVLVA AVSLWATDYGMKSIVGEIVR EIGQKCPQELSRDPSGTKGF AAFLTELAERVPAILMSSMC ILLDHLDGENYMMRNAVLAA MAEMVLQVLSGDQLEAAARD TRDQFLDTLQAHGHDVNSFV RSRVLQLFTRIVQQKALPLT RFQAVVALAVGRLADKSVLV CKNAIQLLASFLANNPFSCK LSDADLAGPLQKETQKLQEM RAQRRTAAASAVLDPEEEWE AMLPELKSTLQQLLQLPQGE EEIPEQIANTETTEDVKGRI YQLLAKASYKKAIILTREAT GHFQESEPFSHIDPEESEET RLLNILGLIFKGPAASTQEK NPRESTGNMVTGQTVCKNKP NMSDPEESRGNDELVKQEML VQYLQDAYSFSRKITEAIGI ISKMMYENTTTVVQEVIEFF VMVFQFGVPQALFGVRRMLP LIWSKEPGVREAVLNAYRQL YLNPKGDSARAKAQALIQNL SLLLVDASVGTIQCLEEILC EFVQKDELKPAVTQLLWERA TEKVACCPLERCSSVMLLGM MARGKPEIVGSNLDTLVSIG LDEKFPQDYRLAQQVCHAIA NISDRRKPSLGKRHPPFRLP QEHRLFERLRETVTKGFVHP DPLWIPFKEVAVTLIYQLAE GPEVICAQILQGCAKQALEK LEEKRTSQEDPKESPAMLPT FLLMNLLSLAGDVALQQLVH LEQAVSGELCRRRVLREEQE HKTKDPKEKNTSSETTMEEE LGLVGATADDTEAELIRGIC EMELLDGKQTLAAFVPLLLK VCNNPGLYSNPDLSAAASLA LGKFCMISATFCDSQLRLLF TMLEKSPLPIVRSNLMVATG DLAIRFPNLVDPWTPHLYAR LRDPAQQVRKTAGLVMTHLI LKDMVKVKGQVSEMAVLLID PEPQIAALAKNFFNELSHKG NAIYNLLPDIISRLSDPELG VEEEPFHTIMKQLLSYITKD KQTESLVEKLCQRFRTSRTE RQQRDLAYCVSQLPLTERGL RKMLDNFDCFGDKLSDESIF SAFLSVVGKLRRGAKPEGKA IIDEFEQKLRACHTRGLDGI KELEIGQAGSQRAPSAKKPS TGSRYQPLASTASDNDFVTP EPRRTTRRHPNTQQRASKKK PKVVFSSDESSEEDLSAEMT EDETPKKTTPILRASARRHR S CapG Homo sapiens Trichoplusia ni LEVO MGAERRLLSIKEAFRLAQQP HQNQAKLVVALSRTYRTMDD KTVFHEEFIHYLKYVMVVYK REPAVERVIEFAAKFVTSFH QSDMEDDEEEEDGGLLNYLF TFLLKSHEANSNAVRFRVCL LINKLLGSMPENAQIDDDVF DKINKAMLIRLKDKIPNVRI QAVLALSRLQDPKDDECPVV NAYATLIENDSNPEVRRAVL SCIAPSAKTLPKIVGRTKDV KEAVRKLAYQVLAEKVHMRA MSIAQRVMLLQQGLNDRSDA VKQAMQKHLLQGWLRFSEGN ILELLHRLDVENSSEVAVSV LNALFSITPLSELVGLCKNN DGRKLIPVETLTPEIALYWC ALCEYLKSKGDEGEEFLEQI LPEPVVYADYLLSYIQSIPV VNEEHRGDFSYIGNLMTKEF IGQQLILIIKSLDTSEEGGR KKLLAVLQEILILPTIPISL VSFLVERLLHIIIDDNKRTQ IVTEIISEIRAPIVTVGVNN DPADVRKKELKMAEIKVKLI EAKEALENCITLQDFNRASE LKEEIKALEDARINLLKETE QLEIKEVHIEKNDAETLQKC LILCYELLKQMSISTGLSAT MNGIIESLILPGIISIHPVV RNLAVLCLGCCGLQNQDFAR KHFVLLLQVLQIDDVTIKIS ALKAIFDQLMTFGIEPFKTK KIKTLHCEGTEINSDDEQES KEVEETATAKNVLKLLSDFL DSEVSELRTGAAEGLAKLMF SGLLVSSRILSRLILLWYNP VTEEDVQLRHCLGVFFPVFA YASRTNQECFEEAFLPTLQT LANAPASSPLAEIDITNVAE LLVDLTRPSGLNPQAKTSQD YQALTVHDNLAMKICNEILT SPCSPEIRVYTKALSSLELS SHLAKDLLVLLNEILEQVKD RTCLRALEKIKIQLEKGNKE FGDQAEAAQDATLTTTTFQN EDEKNKEVYMTPLRGVKATQ ASKSTQLKTNRGQRKVTVSA RTNRRCQTAEADSESDHEVP EPESEMKMRLPRRAKTAALE KSKLNLAQFLNEDLS singleParticle particle 8 20.0 C8H18N2O4S Hepes 200.0 KCl Potassium Chloride

Grafix crosslinked sample was gel filtered on a superose 6 column into EM buffer (20 mM HEPES [pH 8], 200 mM KCl)

NEGATIVE Uranyl Acetate

The grids were glow discharged for 1 minute at 15 mA, the sample applied for 10 seconds and then washed twice with water before staining with 2% uranyl acetate for 1 minute before blotting and drying.

Quantifoil R1.2/1.3 CARBON CONTINUOUS GLOW DISCHARGE FEI TECNAI F20 FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 200 50000.0 SIDE ENTRY, EUCENTRIC TVIPS TEMCAM-F416 (4k x 4k) 4096 4096 15.6 1 862 1.0 50.0 1 12615 CTFFIND 4

RELION initial model

1 C1 BACK PROJECTION 31.8 FSC 0.143 CUT-OFF RELION 3.0 4714 MAXIMUM LIKELIHOOD RELION 3.0 MAXIMUM LIKELIHOOD RELION 3.0 1 4714 RELION 3.0