Poliovirus protein 3AB displays nucleic acid chaperone and helix-destabilizing activities.
Poliovirus protein 3AB displayed nucleic acid chaperone activity in promoting the hybridization of complementary nucleic acids and destabilizing secondary structure. Hybridization reactions at 30 degrees C between 20- and 40-nucleotide RNA oligonucleotides and 179- or 765-nucleotide RNAs that contained a complementary region were greatly enhanced in the presence of 3AB. The effect was nonspecific as reactions between DNA oligonucleotides and RNA or DNA templates were also enhanced. Reactions were optimal with 1 mM MgCl(2) and 20 mM KCl. Analysis of the reactions with various 3AB and template concentrations indicated that enhancement required a critical amount of 3AB that increased as the concentration of nucleic acid increased. This was consistent with a requirement for 3AB to "coat" the nucleic acids for enhancement. The helix-destabilizing activity of 3AB was tested in an assay with two 42-nucleotide completely complementary DNAs. Each complement formed a strong stem-loop (DeltaG = -7.2 kcal/mol) that required unwinding for hybridization to occur. DNAs were modified at the 3' or 5' end with fluorescent probes such that hybridization resulted in quenching of the fluorescent signal. Under optimal conditions at 30 degrees C, 3AB stimulated hybridization in a concentration-dependent manner, as did human immunodeficiency virus nucleocapsid protein, an established chaperone. The results are discussed with respect to the role of 3AB in viral replication and recombination.
DeStefano
Jeffrey J
JJ
Department of Cell Biology and Molecular Genetics, University of Maryland-College Park, Building 231, College Park, MD 20742, USA. jdestefa@umd.edu
Titilope
Oduyebo
O
D003067:Coenzymes; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004795:Enzyme Stability; D018832:Molecular Chaperones; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000378:metabolism; D017955:Poliovirus / Q000502:physiology*; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D014764:Viral Proteins / Q000502:physiology*
D003067:Coenzymes; D018832:Molecular Chaperones; D009841:Oligonucleotides; D014764:Viral Proteins; D012313:RNA; D004247:DNA