Examination of specific binding activity of aptamer RNAs to the HIV-NC by using a cell-based in vivo assay for protein-RNA interaction.
The nucleocapsid (NC) protein of the Human Immunodeficiency Virus-1 plays a key role in viral genomic packaging by specifically recognizing the Psi(Psi) RNA sequence within the HIV-1 genome RNA. Recently, a novel cell-based assay was developed to probe the specific interactions in vivo between the NC and Psi-RNA using E.coli cells (J. Virol. 81: 6151-55, 2007). In order to examine the extendibility of this cell-based assay to RNAs other than Psi-RNA, this study tested the RNA aptamers isolated in vitro using the SELEX method, but whose specific binding ability to NC in a living cellular environment has not been established. The results demonstrate for the first time that each of those aptamer RNAs can bind specifically to NC in a NC zinc finger motif dependent manner within the cell. This confirms that the cell-based assay developed for NC-Psi interaction can be further extended and applied to NC-binding RNAs other than Psi-RNA.
Jeong
Yu Young
YY
National Research Laboratory for Molecular Virology, Department of Pathology, College of Medicine, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul 137-701, Korea.
Kim
Seon Hee
SH
Jang
Soo In
SI
You
Ji Chang
JC
D000595:Amino Acid Sequence; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D001483:Base Sequence; D019411:Clinical Laboratory Techniques; D015497:HIV-1 / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000378:metabolism*; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000032:analysis / Q000378:metabolism; D052156:SELEX Aptamer Technique; D016335:Zinc Fingers; D001616:beta-Galactosidase / Q000378:metabolism
D052157:Aptamers, Nucleotide; D016601:RNA-Binding Proteins; D012313:RNA; D001616:beta-Galactosidase