Novel RNA substrates for the ribozyme from Bacillus subtilis ribonuclease P identified by in vitro selection

Biochemistry
T Pan

Abstract

Novel RNA substrates for the ribozyme from Bacillus subtilis ribonuclease P (P RNA) have been obtained by in vitro selection. The selection method involves cleavage of a circular RNA library by the P RNA, isolation of the linear cleavage product, and regeneration of circular RNA to allow amplification and multiple cycles of selection. The use of circular RNA ensures that potential substrates can be selected without restricted location of the cleavage site. Such a selection method has been used previously to isolate RNA motifs that undergo autolytic cleavage with Pb2+ [Pan, T., & Uhlenbeck, O. (1992) Biochemistry 31, 3887-3895]. The circular RNA pool after eight cycles of selection was cleaved by the B. subtilis P RNA as efficiently as a pre-tRNA(Phe) substrate, estimated to be more than 10 orders of magnitude better than the unselected RNA library. Kinetic analysis of individual variants showed that the kcat/KM of the selected RNA was up to 4-fold higher than that of the pre-tRNA(Phe). When cleavage was carried out with Escherichia coli P RNA, the selected RNA was 10-60-fold less reactive than the reaction of the pre-tRNA(Phe). Two distinct classes of variants are selected, both of which appear to differ significantly from the ...Continue Reading

References

Apr 28, 1992·Biochemistry·T Pan, O C Uhlenbeck
Aug 13, 1992·Nature·T Pan, O C Uhlenbeck
Nov 8, 1991·Science·E S HaasN R Pace
Nov 29, 1991·Science·T PanO C Uhlenbeck
Jan 1, 1990·Annual Review of Biochemistry·T R Cech
Oct 23, 1987·Science·W H McClainS Altman
Nov 11, 1987·Nucleic Acids Research·J F MilliganO C Uhlenbeck
Jan 1, 1980·Methods in Enzymology·T E EnglandO C Uhlenbeck
May 24, 1994·Proceedings of the National Academy of Sciences of the United States of America·E Westhof, S Altman
Oct 11, 1994·Nucleic Acids Research·B K Oh, N R Pace
Sep 27, 1994·Proceedings of the National Academy of Sciences of the United States of America·Y KomineH Inokuchi
Jan 1, 1993·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·S AltmanS Talbot
May 25, 1993·Biochemistry·D Smith, N R Pace

❮ Previous
Next ❯

Citations

Aug 1, 1996·Current Opinion in Biotechnology·R R Breaker
Oct 13, 2007·Quarterly Reviews of Biophysics·Nathan J BairdTobin R Sosnick
Jun 29, 2000·Annual Review of Biochemistry·D S Wilson, J W Szostak
Dec 22, 2010·RNA·Olga EsakovaAndrey S Krasilnikov
Dec 1, 2001·Biological Chemistry·O Gimple, A Schön
May 10, 2013·Annual Review of Biophysics·Alfonso Mondragón
Apr 23, 2008·Current Protocols in Nucleic Acid Chemistry·R R Breaker
Mar 1, 2003·Nature·Andrey S KrasilnikovAlfonso Mondragón
Jul 13, 2020·Trends in Biotechnology·Samara Martín-AlonsoLuis Menéndez-Arias

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.