1161760_Ramakrishnan,S_2021.pdf (4.13 MB)
Synchronous RNA conformational changes trigger ordered phase transitions in crystals
journal contributionposted on 2021-04-13, 03:53 authored by S Ramakrishnan, JR Stagno, CE Conrad, J Ding, P Yu, YR Bhandari, YT Lee, G Pauly, O Yefanov, MO Wiedorn, J Knoska, D Oberthür, TA White, A Barty, V Mariani, C Li, W Brehm, WF Heinz, V Magidson, S Lockett, MS Hunter, S Boutet, Nadia ZatsepinNadia Zatsepin, X Zuo, TD Grant, S Pandey, M Schmidt, JCH Spence, HN Chapman, YX Wang
Time-resolved studies of biomacromolecular crystals have been limited to systems involving only minute conformational changes within the same lattice. Ligand-induced changes greater than several angstroms, however, are likely to result in solid-solid phase transitions, which require a detailed understanding of the mechanistic interplay between conformational and lattice transitions. Here we report the synchronous behavior of the adenine riboswitch aptamer RNA in crystal during ligand-triggered isothermal phase transitions. Direct visualization using polarized video microscopy and atomic force microscopy shows that the RNA molecules undergo cooperative rearrangements that maintain lattice order, whose cell parameters change distinctly as a function of time. The bulk lattice order throughout the transition is further supported by time-resolved diffraction data from crystals using an X-ray free electron laser. The synchronous molecular rearrangements in crystal provide the physical basis for studying large conformational changes using time-resolved crystallography and micro/nanocrystals.
This work was supported in part by the Intramural Research Program of the National Cancer Institute, National Institutes of Health (Y.-X.W.), the NSF-STC "BioXFEL" (NSF-1231306) (J.C.H.S.), the Maxwell computational resources operated at Deutsches Elektronen-Synchrotron (DESY) (H.N.C.), Hamburg, Germany (H.N.C.), and NSF Award (NSF-1565180) (N.A.Z.). Use of the Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, is supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515 (S.B.). Parts of the sample injector used at LCLS for this research were funded by the National Institutes of Health, P41GM103393, formerly P41RR001209.
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