2024-03-20 REL RCSB RCSB RCSB 2019-02-20 2019-03-13 2019-03-13 2024-03-20 Wellcome Trust 200873/Z/16/Z United Kingdom Architectural principles for Hfq/Crc-mediated regulation of gene expression Hfq-Crc-amiE 2:3:2 complex Pei XY Dendooven T Hfq, Crc, amiE, Carbon catabolite repression, RNA-protein interaction, RNA-mediated gene regulation, RNA BINDING PROTEIN, RNA BINDING PROTEIN-RNA-HYDROLASE complex Pei XY Dendooven T Sonnleitner E Chen S Blasi U Luisi BF Architectural principles for Hfq/Crc-mediated regulation of gene expression. Elife US 8 2019 30758287 doi:10.7554/eLife.43158 2050-084X 6o1l FULLOVERLAP Assembly of Hfq-Crc-amiE with a 2:3:2 stoichiometry Assembly of Hfq-Crc-amiE with a 2:3:2 stoichiometry 0 1 2 3 Pseudomonas aeruginosa 0.222 Catabolite repression control protein Pseudomonas aeruginosa 0.030101868999999996 3 Escherichia coli LEVO GPAMRIISVNVNGIQAAAERGLLSWLQAQNADVICLQDTRASAFDLDDPSFQLDGYFLYACDAELPEQGGVALYSRLQPK AVISGLGFETADRYGRYLQADFDKVSIATLLLPSGQSGDESLNQKFKFMDDFTHYLSKQRRKRREYIYCGSLYVAHQKMD VKNWRECQQMPGFLAPERAWLDEVFGNLGYADALREVSREGDQFSWWPDSEQAEMLNLGWRFDYQVLTPGLRRFVRNAKL PRQPRFSQHAPLIVDYDWQLSI Q51380 3.1.11.2 RNA-binding protein Hfq Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1 0.007685984 12 Escherichia coli LEVO HSLQDPYLNTLRKERVPVSIYLVNGIKLQGQIESFDQFVILLKNTVSQMVYKHAISTVVPSRPVRLP Q9HUM0 RNA (5'-R(*AP*AP*AP*AP*AP*UP*AP*AP*CP*AP*AP*CP*AP*AP*GP*AP*GP*G)-3') Pseudomonas aeruginosa 0.00585766 2 AAAAAUAACAACAAGAGG singleParticle particle 7.9 20.0 HEPES pH 7.9 40.0 NaCl 10.0 KCl 1.0 MgCl2 2.0 TCEP Quantifoil, UltrAuFoil, R1.2/1.3 GRAPHENE OXIDE 300 GRAPHENE OXIDE CONTINUOUS GLOW DISCHARGE ETHANE 100 277
Monodisperse
 FEI TITAN KRIOS OTHER BRIGHT FIELD FIELD EMISSION GUN 300 50.0 2.7 0.00125 0.003 125000.0 130841.0 FEI TITAN KRIOS AUTOGRID HOLDER NITROGEN 70.0 100.0 FEI FALCON III (4k x 4k) COUNTING 1 917 60.0 28.0
Images were collected in movie-counting mode, 75 frames per movie stack, 60s exposure per movie
 1 435000 ab-initio map generated in Relion3 C1 3.37 FSC 0.143 CUT-OFF RELION 3b 45311 OTHER RELION 3b beta version
Stochastic Gradient Descent
 MAXIMUM LIKELIHOOD RELION 3b beta version 6 35000.0 RELION 3b beta version emd_0591.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 190 190 190 0 0 0 190 190 190 207.1 207.1 207.1 90.0 90.0 90.0 X Y Z -0.01850905 1.9583755 0.0037314594 0.054603253 1.09 1.09 1.09 0.27 AUTHOR Locally sharpened Cryo-EM map of a Hfq-Crc-amiE assembly with stoichiometry 2:3:2, respectively. 1U1T PDB experimental model 4JG3 PDB experimental model RIGID BODY FIT Cross-correlation REAL emd_0591_msk_1.map emd_0591_additional_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 190 190 190 0 0 0 190 190 190 207.1 207.1 207.1 90.0 90.0 90.0 X Y Z -0.1668402 0.23806459 0.000016997947 0.012787011 1.09 1.09 1.09 AUTHOR Globally sharpened Cryo-EM map of a Hfq-Crc-amiE assembly with stoichiometry 2:3:2, respectively. emd_0591_half_map_1.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 190 190 190 0 0 0 190 190 190 207.1 207.1 207.1 90.0 90.0 90.0 X Y Z -0.06508681 0.11658159 0.000015071115 0.0077043055 1.09 1.09 1.09 AUTHOR Half map 1 emd_0591_half_map_2.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 190 190 190 0 0 0 190 190 190 207.1 207.1 207.1 90.0 90.0 90.0 X Y Z -0.058557525 0.11448459 0.00001906727 0.0076821907 1.09 1.09 1.09 AUTHOR Half map 2