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Biochemistry. 2014 Jul 15. [Epub ahead of print]
Secondary Structure of a Conserved Domain in an Intron of Influenza A M1 mRNA.
Jiang T, Kennedy SD, Moss WN, Kierzek E, Turner DH.
Abstract
Influenza A virus utilizes RNA throughout infection. Little is known, however, about the roles of RNA structures in these processes. A previous bioinformatics survey predicted multiple regions of influenza A virus that are likely to generate evolutionarily conserved and stable RNA structures. One predicted conserved structure is in the pre-mRNA coding for essential proteins, M1 and M2. This structure is 39 nucleotides downstream of the M2 mRNA 5' splice site. Here a combination of biochemical structural mapping, mutagenesis, and NMR confirms the predicted three-way multi-branch structure of this RNA. Imino proton spectra reveal no change in secondary structure when 80 mM KCl is supplemented with 4 mM MgCl2. Optical melting curves in 1 M NaCl and in 100 mM KCl with 10 mM MgCl2 are very similar, with melting temperatures ~14 ?C higher than for 100 mM KCl alone. These results provide a firm basis for designing experiments and potential therapeutics to test for function in cell culture.
PMID:
25026548
[PubMed - as supplied by publisher]
Secondary Structure of a Conserved Domain in an Intron of Influenza A M1 mRNA.
Jiang T, Kennedy SD, Moss WN, Kierzek E, Turner DH.
Abstract
Influenza A virus utilizes RNA throughout infection. Little is known, however, about the roles of RNA structures in these processes. A previous bioinformatics survey predicted multiple regions of influenza A virus that are likely to generate evolutionarily conserved and stable RNA structures. One predicted conserved structure is in the pre-mRNA coding for essential proteins, M1 and M2. This structure is 39 nucleotides downstream of the M2 mRNA 5' splice site. Here a combination of biochemical structural mapping, mutagenesis, and NMR confirms the predicted three-way multi-branch structure of this RNA. Imino proton spectra reveal no change in secondary structure when 80 mM KCl is supplemented with 4 mM MgCl2. Optical melting curves in 1 M NaCl and in 100 mM KCl with 10 mM MgCl2 are very similar, with melting temperatures ~14 ?C higher than for 100 mM KCl alone. These results provide a firm basis for designing experiments and potential therapeutics to test for function in cell culture.
PMID:
25026548
[PubMed - as supplied by publisher]
