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- EMDB-10912: Cryo-EM structure of T7 bacteriophage DNA translocation gp15-gp16... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-10912 | ||||||||||||
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Title | Cryo-EM structure of T7 bacteriophage DNA translocation gp15-gp16 core complex intermediate assembly | ||||||||||||
![]() | t7 tail complex | ||||||||||||
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![]() | VIRAL COMPLEX / DNA TRANSLOCATION / VIRAL INFECTION / TRANS-GLYCOSYLASE ACTIVITY / CHAPERONE / PERIPLASMIC SPACE COMPLEX / VIRAL PROTEIN | ||||||||||||
Function / homology | ![]() symbiont genome ejection through host cell envelope / host cell periplasmic space / : / symbiont entry into host cell via disruption of host cell wall peptidoglycan / lytic transglycosylase activity / symbiont genome ejection through host cell envelope, short tail mechanism / symbiont entry into host cell via disruption of host cell envelope / peptidoglycan metabolic process / symbiont entry into host / virion component ...symbiont genome ejection through host cell envelope / host cell periplasmic space / : / symbiont entry into host cell via disruption of host cell wall peptidoglycan / lytic transglycosylase activity / symbiont genome ejection through host cell envelope, short tail mechanism / symbiont entry into host cell via disruption of host cell envelope / peptidoglycan metabolic process / symbiont entry into host / virion component / killing of cells of another organism / defense response to bacterium / hydrolase activity / host cell plasma membrane / membrane Similarity search - Function | ||||||||||||
Biological species | ![]() ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | ||||||||||||
![]() | Perez-Ruiz M / Pulido-Cid M | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Assisted assembly of bacteriophage T7 core components for genome translocation across the bacterial envelope. Authors: Mar Pérez-Ruiz / Mar Pulido-Cid / Juan Román Luque-Ortega / José María Valpuesta / Ana Cuervo / José L Carrascosa / ![]() Abstract: In most bacteriophages, genome transport across bacterial envelopes is carried out by the tail machinery. In viruses of the family, in which the tail is not long enough to traverse the bacterial ...In most bacteriophages, genome transport across bacterial envelopes is carried out by the tail machinery. In viruses of the family, in which the tail is not long enough to traverse the bacterial wall, it has been postulated that viral core proteins assembled inside the viral head are translocated and reassembled into a tube within the periplasm that extends the tail channel. Bacteriophage T7 infects , and despite extensive studies, the precise mechanism by which its genome is translocated remains unknown. Using cryo-electron microscopy, we have resolved the structure of two different assemblies of the T7 DNA translocation complex composed of the core proteins gp15 and gp16. Gp15 alone forms a partially folded hexamer, which is further assembled upon interaction with gp16 into a tubular structure, forming a channel that could allow DNA passage. The structure of the gp15-gp16 complex also shows the location within gp16 of a canonical transglycosylase motif involved in the degradation of the bacterial peptidoglycan layer. This complex docks well in the tail extension structure found in the periplasm of T7-infected bacteria and matches the sixfold symmetry of the phage tail. In such cases, gp15 and gp16 that are initially present in the T7 capsid eightfold-symmetric core would change their oligomeric state upon reassembly in the periplasm. Altogether, these results allow us to propose a model for the assembly of the core translocation complex in the periplasm, which furthers understanding of the molecular mechanism involved in the release of T7 viral DNA into the bacterial cytoplasm. | ||||||||||||
History |
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Structure visualization
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 111.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 17.1 KB 17.1 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.4 KB | Display | ![]() |
Images | ![]() | 70.3 KB | ||
Filedesc metadata | ![]() | 7.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6yt5MC ![]() 6yszC C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | t7 tail complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.047 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : gp15-gp16 core complex
Entire | Name: gp15-gp16 core complex |
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Components |
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-Supramolecule #1: gp15-gp16 core complex
Supramolecule | Name: gp15-gp16 core complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 1.4 MDa |
-Macromolecule #1: Internal virion protein gp15
Macromolecule | Name: Internal virion protein gp15 / type: protein_or_peptide / ID: 1 / Number of copies: 6 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 88.377219 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDPSSMSKIE SALQAAQPGL SRLRGGAGGM GYRAATTQAE QPRSSLLDTI GRFAKAGAD MYTAKEQRAR DLADERSNEI IRKLTPEQRR EALNNGTLLY QDDPYAMEAL RVKTGRNAAY LVDDDVMQKI K EGVFRTRE ...String: MRGSHHHHHH GMASMTGGQQ MGRDLYDDDD KDPSSMSKIE SALQAAQPGL SRLRGGAGGM GYRAATTQAE QPRSSLLDTI GRFAKAGAD MYTAKEQRAR DLADERSNEI IRKLTPEQRR EALNNGTLLY QDDPYAMEAL RVKTGRNAAY LVDDDVMQKI K EGVFRTRE EMEEYRHSRL QEGAKVYAEQ FGIDPEDVDY QRGFNGDITE RNISLYGAHD NFLSQQAQKG AIMNSRVELN GV LQDPDML RRPDSADFFE KYIDNGLVTG AIPSDAQATQ LISQAFSDAS SRAGGADFLM RVGDKKVTLN GATTTYRELI GEE QWNALM VTAQRSQFET DAKLNEQYRL KINSALNQED PRTAWEMLQG IKAELDKVQP DEQMTPQREW LISAQEQVQN QMNA WTKAQ AKALDDSMKS MNKLDVIDKQ FQKRINGEWV STDFKDMPVN ENTGEFKHSD MVNYANKKLA EIDSMDIPDG AKDAM KLKY LQADSKDGAF RTAIGTMVTD AGQEWSAAVI NGKLPERTPA MDALRRIRNA DPQLIAALYP DQAELFLTMD MMDKQG IDP QVILDADRLT VKRSKEQRFE DDKAFESALN ASKAPEIARM PASLRESARK IYDSVKYRSG NESMAMEQMT KFLKEST YT FTGDDVDGDT VGVIPKNMMQ VNSDPKSWEQ GRDILEEARK GIIASNPWIT NKQLTMYSQG DSIYLMDTTG QVRVRYDK E LLSKVWSENQ KKLEEKAREK ALADVNKRAP IVAATKAREA AAKRVREKRK QTPKFIYGRK E UniProtKB: Internal virion protein gp15 |
-Macromolecule #2: Peptidoglycan transglycosylase gp16
Macromolecule | Name: Peptidoglycan transglycosylase gp16 / type: protein_or_peptide / ID: 2 / Number of copies: 6 / Enantiomer: LEVO EC number: Lyases; Carbon-oxygen lyases; Acting on polysaccharides |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 146.693094 KDa |
Recombinant expression | Organism: ![]() ![]() |
Sequence | String: MGHHHHHHHH HHSSGHIEGR HMMDKYDKNV PSDYDGLFQK AADANGVSYD LLRKVAWTES RFVPTAKSKT GPLGMMQFTK ATAKALGLR VTDGPDDDRL NPELAINAAA KQLAGLVGKF DGDELKAALA YNQGEGRLGN PQLEAYSKGD FASISEEGRN Y MRNLLDVA ...String: MGHHHHHHHH HHSSGHIEGR HMMDKYDKNV PSDYDGLFQK AADANGVSYD LLRKVAWTES RFVPTAKSKT GPLGMMQFTK ATAKALGLR VTDGPDDDRL NPELAINAAA KQLAGLVGKF DGDELKAALA YNQGEGRLGN PQLEAYSKGD FASISEEGRN Y MRNLLDVA KSPMAGQLET FGGITPKGKG IPAEVGLAGI GHKQKVTQEL PESTSFDVKG IEQEATAKPF AKDFWETHGE TL DEYNSRS TFFGFKNAAE AELSNSVAGM AFRAGRLDNG FDVFKDTITP TRWNSHIWTP EELEKIRTEV KNPAYINVVT GGS PENLDD LIKLANENFE NDSRAAEAGL GAKLSAGIIG AGVDPLSYVP MVGVTGKGFK LINKALVVGA ESAALNVASE GLRT SVAGG DADYAGAALG GFVFGAGMSA ISDAVAAGLK RSKPEAEFDN EFIGPMMRLE ARETARNANS ADLSRMNTEN MKFEG EHNG VPYEDLPTER GAVVLHDGSV LSASNPINPK TLKEFSEVDP EKAARGIKLA GFTEIGLKTL GSDDADIRRV AIDLVR SPT GMQSGASGKF GATASDIHER LHGTDQRTYN DLYKAMSDAM KDPEFSTGGA KMSREETRYT IYRRAALAIE RPELQKA LT PSERIVMDII KRHFDTKREL MENPAIFGNT KAVSIFPESR HKGTYVPHVY DRHAKALMIQ RYGAEGLQEG IARSWMNS Y VSRPEVKARV DEMLKELHGV KEVTPEMVEK YAMDKAYGIS HSDQFTNSSI IEENIEGLVG IENNSFLEAR NLFDSDLSI TMPDGQQFSV NDLRDFDMFR IMPAYDRRVN GDIAIMGSTG KTTKELKDEI LALKAKAEGD GKKTGEVHAL MDTVKILTGR ARRNQDTVW ETSLRAINDL GFFAKNAYMG AQNITEIAGM IVTGNVRALG HGIPILRDTL YKSKPVSAKE LKELHASLFG K EVDQLIRP KRADIVQRLR EATDTGPAVA NIVGTLKYST QELAARSPWT KLLNGTTNYL LDAARQGMLG DVISATLTGK TT RWEKEGF LRGASVTPEQ MAGIKSLIKE HMVRGEDGKF TVKDKQAFSM DPRAMDLWRL ADKVADEAML RPHKVSLQDS HAF GALGKM VMQFKSFTIK SLNSKFLRTF YDGYKNNRAI DAALSIITSM GLAGGFYAMA AHVKAYALPK EKRKEYLERA LDPT MIAHA ALSRSSQLGA PLAMVDLVGG VLGFESSKMA RSTILPKDTV KERDPNKPYT SREVMGAMGS NLLEQMPSAG FVANV GATL MNAAGVVNSP NKATEQDFMT GLMNSTKELV PNDPLTQQLV LKIYEANGVN LRERRK UniProtKB: Peptidoglycan transglycosylase gp16 |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 1.5 mg/mL | ||||||||
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Buffer | pH: 8 Component:
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Grid | Model: Quantifoil R2/2 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec. / Pretreatment - Atmosphere: AIR | ||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 293.15 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 2 / Number real images: 5506 / Average exposure time: 8.0 sec. / Average electron dose: 40.8 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 47619 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen 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
-Atomic model buiding 1
Initial model |
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Refinement | Space: REAL / Protocol: AB INITIO MODEL | ||||||
Output model | ![]() PDB-6yt5: |