RNA-binding properties of the matrix protein (p19gag) of avian sarcoma and leukemia viruses.
We have reinvestigated the ability of the matrix protein (MA) (p19gag) of avian sarcoma and leukemia viruses to interact with RNA. Previous reports claimed on the one hand that MA can bind tightly and with a high degree of specificity to avian sarcoma and leukemia virus RNA in vitro and on the other that it cannot bind to RNA at all. We found that MA purified by any of several methods does bind to RNA, as measured by its ability to cause retention of radioactive RNA on nitrocellulose membranes in a filtration assay. However, this interaction is weak and lacks specificity. The interaction of MA with RNA was barely detectable by classical sedimentation analysis, and from this observation we estimate that the intrinsic MA-RNA association constant is ca. 10(3) M-1, at least 3 orders of magnitude smaller than the constant describing the interaction of the viral nucleocapsid protein (NC) (p12gag) with RNA, ca. 10(6) M-1. Separately purified phosphorylated and nonphosphorylated MA species bound RNA equally. We also found that MA can bind to DNA with an affinity similar to that for RNA. The large quantitative discrepancy between our results and earlier published reports can be traced in part to methods of data analysis.
Steeg
C M
CM
Section of Biochemistry and Molecular and Cellular Biology, Cornell University, Ithaca, New York 14853.
Vogt
V M
VM
D001354:Avian Leukosis Virus / Q000378:metabolism*; D009189:Avian Myeloblastosis Virus / Q000378:metabolism*; D001358:Avian Sarcoma Viruses / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015683:Gene Products, gag / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D010766:Phosphorylation; D010957:Plasmids; D011485:Protein Binding; D012335:RNA, Ribosomal / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D014158:Transcription, Genetic
D004279:DNA, Viral; D015683:Gene Products, gag; D012335:RNA, Ribosomal; D012367:RNA, Viral