tailieunhanh - Báo cáo khoa học: Dissecting the role of protein–protein and protein–nucleic acid interactions in MS2 bacteriophage stability

To investigate the role of protein–protein and protein–nucleic acid interac-tions in virus assembly, we compared the stabilities of native bacteriophage MS2, virus-like particles (VLPs) containing nonviral RNAs, and an assem-bly-defective coat protein mutant (dlFG) and its single-chain variant (sc-dlFG). | ềFEBS Journal Dissecting the role of protein-protein and protein-nucleic acid interactions in MS2 bacteriophage stability Sheila M. B. Lima1 Ana Carolina Q. Vaz1 Theo L. F. Souza1 David S. Peabody2 Jerson L. Silva1 and Andrea C. Oliveira1 1 Programa de Biologia Estruturaland Centro Nacionalde Ressonancia Magnetica Nuclear de Macromoleculas Institute de Bioquimica Medica Universidade Federaldo Rio de Janeiro Brazil 2 Department of Molecular Genetics and Microbiology and Cancer Research and Treatment Center University of New Mexico Schoolof Medicine Albuquerque NM USA Keywords coat protein interactions fluorescence hydrostatic pressure MS2 bacteriophage virus-like particles Correspondence J. L. Silva and A. C. Oliveira Avenida Bauhinia 400 - CCS ICB Bl E sl. 08 Cidade Universitaria CEP Rio de Janeiro RJ 21941-590 Brazil Fax 55 21 3881 4155 Tel 55 21 2562 6756 E-mail jerson@ E-mail cheble@ Received 2 August 2005 revised 26 January 2006 accepted 6 February 2006 doi To investigate the role of protein-protein and protein-nucleic acid interactions in virus assembly we compared the stabilities of native bacteriophage MS2 virus-like particles VLPs containing nonviral RNAs and an assembly-defective coat protein mutant dlFG and its single-chain variant sc-dlFG . Physical high pressure and chemical urea and guanidine hydrochloride agents were used to promote virus disassembly and protein denaturation and the changes in virus and protein structure were monitored by measuring tryptophan intrinsic fluorescence bis-ANS probe fluorescence and light scattering. We found that VLPs dissociate into capsid proteins that remain folded and more stable than the proteins dissociated from authentic particles. The proposed model is that the capsid disassembles but the protein remains bound to the heterologous RNA encased by VLPs. The dlFG dimerizes correctly but fails to assemble into capsids because it lacks the 15-amino acid FG

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