In vitro selection of ATP-binding receptors using a ribonucleopeptide complex.
A recently described three-dimensional structure of the ribosome provides a sense of remarkable diversity of RNA-protein complexes. We have designed a new class of scaffold for artificial receptors, in which a short peptide and RNA with a randomized nucleotide region form a stable and specific complex. The randomized nucleotide region was placed next to the HIV-1 Rev response element to enable the formation of "ribonucleopeptide" pools in the presence of the Rev peptide. In vitro selection of RNA oligonucleotides from the randomized pool afforded a ribonucleopeptide receptor specific for ATP. The ATP-binding ribonucleopeptide did not share the known consensus nucleotide sequence for ATP aptamers and completely lost its ATP-binding ability in the absence of the Rev peptide. The ATP-binding activity of the ribonucleopeptide was increased by a substitution of the N-terminal amino acid of the Rev peptide. These results demonstrate directly that the peptide is incorporated in the functional structure of RNA and suggest that amino acids outside the RNA-binding region of the peptide modulate the ATP-binding of ribonucleopeptide. Our approach would provide an alternative strategy for the design of "tailor-made" ribonucleopeptide receptors and enzymes.
Morii
Takashi
T
Institute of Advanced Energy, Kyoto University, and PRESTO, Japan Science and Technology Corporation, Uji, Kyoto 611-0011, Japan. t-morii@iae.kyoto-u.ac.jp
Hagihara
Masaki
M
Sato
Shin-ichi
S
Makino
Keisuke
K
D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D015694:Gene Products, rev / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D018048:Receptors, Purinergic P2 / Q000737:chemistry* / Q000378:metabolism; D020218:Response Elements; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000378:metabolism
D015694:Gene Products, rev; D010446:Peptide Fragments; D018048:Receptors, Purinergic P2; D012261:Ribonucleoproteins; D012313:RNA; D000255:Adenosine Triphosphate