Hammerhead ribozyme cleavage of hamster prion pre-mRNA in complex cell-free model systems.
The cleavage properties of a trans-acting hammerhead ribozyme targeted 51 bases upstream of the putative splicing branch point in the hamster prion pre-mRNA intron were investigated in cell-free model systems in vitro. The specificity of cleavage was demonstrated by the inability of this ribozyme to cleave a non-homologous synthetic message encoding part of the beta amyloid peptide precursor, beta APP, and by the inability of the prion pre-mRNA to be cleaved by a ribozyme targeted to beta amyloid peptide precursor mRNA. Also, the addition of total RNA isolated from rat brain had only a minimal effect on the cleavage of the prion substrate pre-mRNA by the ribozyme. Finally neither the presence of 100 ng of nuclear or cytoplasmic proteins were found to affect the rate of cleavage in vitro.
Denman
R B
RB
New York State Institute for Basic Research in Developmental Disabilities, Staten Island 10314.
Purow
B
B
Rubenstein
R
R
Miller
D L
DL
D000255:Adenosine Triphosphate / Q000378:metabolism; D016564:Amyloid beta-Protein Precursor / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000502:physiology; D002474:Cell-Free System; D003001:Cloning, Molecular; D006224:Cricetinae; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011328:Prions / Q000235:genetics*; D012313:RNA / Q000235:genetics; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism*; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D015183:Restriction Mapping; D012754:Sharks; D013379:Substrate Specificity; D014158:Transcription, Genetic
D016564:Amyloid beta-Protein Precursor; D011328:Prions; D012322:RNA Precursors; D016337:RNA, Catalytic; D012333:RNA, Messenger; D012313:RNA; D000255:Adenosine Triphosphate