rna packing specificity and folding during assembly of the bacteriophage ms2rna特异性和折叠包装在噬菌体的组装一份.pdf

Computational and Mathematical Methods in Medicine Vol. 9, Nos. 3–4, September–December 2008, 339–349 RNA packing specificity and folding during assembly of the bacteriophage MS2 1 Ottar Rolfsson, Katerina Toropova, Victoria Morton, Simona Francese , Gabriella Basnak, Gary S. Thompson, Stephen W. Homans, Alison E. Ashcroft, Nicola J. Stonehouse, Neil A. Ranson and Peter G. Stockley* Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK ( Received 18 February 2008; final version received 1 March 2008 ) Using a combination of biochemistry, mass spectrometry, NMR spectroscopy and cryo- electron microscopy (cryo-EM), we have been able to show that quasi-equivalent conformer switching in the coat protein (CP) of an RNA bacteriophage (MS2) is controlled by a sequence-specific RNA–protein interaction. The RNA component of this complex is an RNA stem-loop encompassing just 19 nts from the phage genomic RNA, which is 3569 nts in length. This binding results in the conversion of a CP dimer from a symmetrical conformation to an asymmetric one. Only when both symmetrical and asymmetrical dimers are present in solution is assembly of the T ¼ 3 phage capsid efficient. This implies that the conformers, we have characterized by NMR correspond to the two distinct quasi-equivalent conformers seen in the 3D structure of the virion. An icosahedrally-averaged single particle cryo-EM ˚ reconstruction of the wild-type phage (to ,9 A resolution) has revealed icosahedrally ordered density encompassing up to 90%