2020-01-29RELRCSBRCSBRCSB2016-10-252016-11-162016-12-142020-01-29Jane Coffin Childs Foundation post-doctoral fellowshipUnited StatesLeona M. and Harry B. Helmsley Charitable Trust Grant2012-PG-MED002United StatesNational Institute of Aging K99 transitional awardAG050749United StatesNational Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM053757United StatesNational Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM103310United StatesAgency for Science, Technology and Research SingaporeSingaporeSimons Foundation349247United StatesStructure of 30S Ribosome - Class FTan YDavis JHLyumkis DDavis JHTan YZCarragher BPotter CSLyumkis DWilliamson JRModular Assembly of the Bacterial Large Ribosomal Subunit.Cell16716101622.e15201627912064doi:10.1016/j.cell.2016.11.0201097-4172EMD-8457associated EM volumeEMD-8447other EM volumeEMD-8446other EM volumeEMD-8445other EM volumeEMD-8444other EM volumeEMD-8443other EM volumeEMD-8442other EM volumeEMD-8441other EM volumeEMD-8440other EM volumeEMD-8452other EM volumeEMD-8451other EM volumeEMD-8450other EM volumeEMD-8449other EM volumeEMD-8448other EM volumeEMD-8453other EM volumeEMD-8455other EM volumeEMD-8456other EM volume30S Ribosome - Class F30S Ribosome - Class F0E. coli small ribosome subunit purified from bL17-depleted cells.Escherichia coliNCM3722 rplQ::cat, pHSL-rplQsingleParticleparticle7.550.0C4H11NO3Tris Hydrochloride100.0NH4ClAmmonium Chloride10.0MgCl2Magnesium Chloride0.5C10H16N2O8Ethylenediaminetetraacetic Acid6.0C2H6OSBeta-mercaptoethanolMost of the sucrose was removed by spin concentrationC-Flat CF-1.2/1.3-4AuGOLD400PLASMA CLEANINGOTHERETHANE80298GATAN CRYOPLUNGE 33 microliter of the sample was added.. Fraction was obtained directly from a sucrose gradient, and was spin-concentrated in a 100 kDa MW-cutoff filter.FEI TITAN KRIOSFLOOD BEAMBRIGHT FIELDFIELD EMISSION GUN300100.02.71.03.522500.038167.0FEI TITAN KRIOS AUTOGRID HOLDERNITROGENIn order to account for highly preferred orientation of the specimen, data was acquired using tilts ranging from 10-60 degrees, in addition to untilted images at 0 degrees. The CTF was estimated on a per-particle basis to account for the gradient of CTF values across individual micrographsGATAN K2 SUMMIT (4k x 4k)COUNTING371038385.01-502247910.034.27144701784028 particles were initially picked with DoG picker. These particles were put through 2D classification. Good class 2D class averages then served as templates for FindEM. FindEM picked a total of 447017 particles.CTFFIND3CTFTILTGctf0.5AppionAb initio model was produced from the 2D class averages using Optimod in the Appion pipeline.1C1FOURIER SPACE7.9FSC 0.143 CUT-OFFRELION1.41853PROJECTION MATCHINGRELION1.4PROJECTION MATCHINGFREALIGN9.0755600RELION1.4emd_8457_msk_1.mapemd_8457_additional.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320419.19998419.19998419.1999890.090.090.0XYZ-0.0376403970.08787246-0.00136735910.0052140151.311.311.31Class F 30S Ribosome - Unsharpenedemd_8457_half_map_1.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320419.19998419.19998419.1999890.090.090.0XYZ-1.0686031.0185025-0.00304372030.075775891.311.311.31Class F 30S Ribosome - Halfmap 2emd_8457_half_map_2.map.gz1IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
320
320320
0
00320320320419.19998419.19998419.1999890.090.090.0XYZ-0.98849270.9991745-0.0025486290.076098131.311.311.31Class F 30S Ribosome - Halfmap 1