Identification of tolerated insertion sites in poliovirus non-structural proteins.
Insertion of nucleotide sequences encoding "tags" that can be expressed in specific viral proteins during an infection is a useful strategy for purifying viral proteins and their functional complexes from infected cells and/or for visualizing the dynamics of their subcellular location over time. To identify regions in the poliovirus polyprotein that could potentially accommodate insertion of tags, transposon-mediated insertion mutagenesis was applied to the entire nonstructural protein-coding region of the poliovirus genome, followed by selection of genomes capable of generating infectious, viable viruses. This procedure allowed us to identify at least one site in each viral nonstructural protein, except protein 2C, in which a minimum of five amino acids could be inserted. The distribution of these sites is analyzed from the perspective of their protein structural context and from the perspective of virus evolution.
Laboratory of Infectious Diseases, NIAID, NIH, Bethesda, MD 20892, USA. nteterina@niaid.nih.gov
Natalya L
NL
Teterina
Chris
C
Lauber
Kenneth S
KS
Jensen
Eric A
EA
Levenson
Alexander E
AE
Gorbalenya
Ellie
E
Ehrenfeld
D000595:Amino Acid Sequence; D001483:Base Sequence; D004251:DNA Transposable Elements / Q000235:genetics*; D019143:Evolution, Molecular; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D017955:Poliovirus / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D004251:DNA Transposable Elements; D017361:Viral Nonstructural Proteins