Protein enhancement of hammerhead ribozyme catalysis.
When the recognition sequence of a ribozyme is extended beyond a certain length, turnover is slowed and specificity is decreased. Here, it is shown that a protein can help a ribozyme overcome these general limitations on ribozyme activity. Cleavage of an RNA oligonucleotide by a hammerhead ribozyme is enhanced 10- to 20-fold upon addition of a protein derived from the p7 nucleocapsid (NC) protein of human immunodeficiency virus-type 1. The NC protein also enhances the ability of the ribozyme to discriminate between cleavage of RNA oligonucleotides with differing sequences. These catalytic improvements can be attributed to the strand exchange activity of this RNA binding protein. It is conceivable that endogenous or added proteins may provide analogous increases in ribozyme activity and specificity in vivo.
Tsuchihashi
Z
Z
Howard Hughes Medical Institute, Stanford University, CA 94305.
Khosla
M
M
Herschlag
D
D
D001483:Base Sequence; D036022:Capsid Proteins; D002384:Catalysis; D004277:DNA, Single-Stranded / Q000378:metabolism; D015683:Gene Products, gag / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009843:Oligoribonucleotides / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D014764:Viral Proteins; D016335:Zinc Fingers; D054301:gag Gene Products, Human Immunodeficiency Virus
D036022:Capsid Proteins; D004277:DNA, Single-Stranded; D015683:Gene Products, gag; C071884:NCP7 protein, Human immunodeficiency virus 1; D009843:Oligoribonucleotides; D016337:RNA, Catalytic; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; D012313:RNA