Nickel2+-mediated assembly of an RNA-amino acid complex.
The RNA species SHR1 reacts with biocytin (epsilon-biotinoyl-L-lysine) in the presence of Ni(2+) or Pt(2+) to produce a metal-bridged complex that migrates more slowly than unreacted RNA in the presence of streptavidin (StrAv) on denaturing polyacrylamide gels. Mapping of reverse transcription pause sites identified G79 as a reactive nucleotide. G79 is near the 3' end of a 37 nucleotide core motif that is nearly as reactive as SHR1. SHR1 reacts with biocytin in the presence of Pt(2+) to yield a product that comigrates with the Ni(2+) product but that is much more stable, suggesting that the metal ion used in the reaction is present in the product, possibly linking the RNA to the amino acid. In support of this model, SHR1 shows a strong affinity for Ni(2+) in immobilized metal ion chromatography.
Hati
Sanchita
S
Department of Chemistry, Indiana University, Bloomington, IN 47405, USA. dhburke@indiana.edu
Boles
Amy R
AR
Zaborske
John M
JM
Bergman
Brett
B
Posto
Amanda L
AL
Burke
Donald H
DH
D000596:Amino Acids / Q000737:chemistry*; D001483:Base Sequence; D017931:DNA Primers; D009532:Nickel / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*
D000596:Amino Acids; D017931:DNA Primers; D012313:RNA; D009532:Nickel