REL OBS RCSB RCSB RCSB 2010-03-09 2010-05-21 2010-09-01 2017-09-27 2017-09-27 Liu H Jin L Koh SBS Atanasov I Schein S Wu L Zhou ZH human adenovirus, cryoEM, 3D reconstruction, full-atom model, interaction network Liu H Jin L Koh SB Atanasov I Schein S Wu L Zhou ZH SCIENCE 329 1038 1043 2010 20798312 doi:10.1126/science.1187433 3iyn FULLOVERLAP human adenovirus type 5 human adenovirus type 5

E1B gene-attenuated oncolytic human adenovirus type 5 was propagated in HEK 293 cells, harvested and liberated by 3 cycles of freezing-thawing and purified by CsCl step gradient ultra-centrifugation. The virion particles were then dialyzed and resuspended in 10 mM Tris (pH 7.5), 1 mM MgCl2.

Icosahedral particle 15 Human adenovirus 5 Human adenovirus 5 Homo sapiens VERTEBRATES 150 150 920 VIRION SEROTYPE false false human adenovirus type 5 singleParticle particle 7.5

10mM Tris-HCL,1mM MgCl2

NEGATIVE

The virus particles were not stained.

200 mesh holey carbon grid

ETHANE 100 90 OTHER

Vitrification instrument: FEI Vitrobot. Vitrification carried out in a chamber with controlled humidity.

Blot for 4 seconds before plunging FEI TITAN KRIOS FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 300 2.7 1.0 2.5 59000.0 OTHER 90 objective lens astigmatism was corrected at 100,000 times magnification 0

low-dose

2009-05-01 KODAK SO-163 FILM NIKON SUPER COOLSCAN 9000 6.35 756 20 8.0 Eucentric 0 0

756 micrographs that clearly showed signals up to 4 Angstrom in their power spectra were selected. Individual particle images (1024 pixel by 1024 pixel) were first boxed out automatically by the autoBox program in the IMIRS package, followed by manual screening with the EMAN boxer program to keep only the well-separated, contamination-free and intact 45000 particles. The program CTFFIND was used to determine the defocus value and astigmatism parameters for each micrograph. Determination of particle orientation and centre, and subsequent 3D reconstruction were done with the IMIRS package, enhanced by icosahedral symmetry-adapted functions.

Each particle

OTHER 3.6 FSC 0.5 CUT-OFF IMIRS

The program CTFFIND was used to determine the defocus value and astigmatism parameters for each micrograph. We determined particle orientation, centre parameters and subsequent 3D reconstruction with the IMIRS package, enhanced by icosahedral symmetry-adapted functions for 3D reconstruction. We incorporated astigmatism in the CTF correction in both orientation-centre refinement and 3D reconstruction steps. The IMIRS procedure was optimized, and the data processing was completed within two months by three personal computers, each with eight 2.33GHz CPUs and 16G memory.

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