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Optimizing the substrate specificity of a group I intron ribozyme.

Publication ,  Journal Article
Zarrinkar, PP; Sullenger, BA
Published in: Biochemistry
March 16, 1999

Group I ribozymes can repair mutant RNAs via trans-splicing. Unfortunately, substrate specificity is quite low for the trans-splicing reaction catalyzed by the group I ribozyme from Tetrahymenathermophila. We have used a systematic approach based on biochemical knowledge of the function of the Tetrahymena ribozyme to optimize its ability to discriminate against nonspecific substrates in vitro. Ribozyme derivatives that combine a mutation which indirectly slows down the rate of the chemical cleavage step by weakening guanosine binding with additional mutations that weaken substrate binding have greatly enhanced specificity with short oligonucleotide substrates and an mRNA fragment derived from the p53 gene. Moreover, compared to the wild-type ribozyme, reaction of a more specific ribozyme with targeted substrates is much less sensitive to the presence of nonspecific RNA competitors. These results demonstrate how a detailed understanding of the biochemistry of a catalytic RNA can facilitate the design of customized ribozymes with improved properties for therapeutic applications.

Duke Scholars

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 16, 1999

Volume

38

Issue

11

Start / End Page

3426 / 3432

Location

United States

Related Subject Headings

  • Tetrahymena thermophila
  • Substrate Specificity
  • RNA, Messenger
  • RNA, Catalytic
  • Nucleic Acid Conformation
  • Molecular Sequence Data
  • Models, Chemical
  • Models, Biological
  • Kinetics
  • Introns
 

Citation

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Zarrinkar, P. P., & Sullenger, B. A. (1999). Optimizing the substrate specificity of a group I intron ribozyme. Biochemistry, 38(11), 3426–3432. https://doi.org/10.1021/bi982688m
Zarrinkar, P. P., and B. A. Sullenger. “Optimizing the substrate specificity of a group I intron ribozyme.Biochemistry 38, no. 11 (March 16, 1999): 3426–32. https://doi.org/10.1021/bi982688m.
Zarrinkar PP, Sullenger BA. Optimizing the substrate specificity of a group I intron ribozyme. Biochemistry. 1999 Mar 16;38(11):3426–32.
Zarrinkar, P. P., and B. A. Sullenger. “Optimizing the substrate specificity of a group I intron ribozyme.Biochemistry, vol. 38, no. 11, Mar. 1999, pp. 3426–32. Pubmed, doi:10.1021/bi982688m.
Zarrinkar PP, Sullenger BA. Optimizing the substrate specificity of a group I intron ribozyme. Biochemistry. 1999 Mar 16;38(11):3426–3432.
Journal cover image

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 16, 1999

Volume

38

Issue

11

Start / End Page

3426 / 3432

Location

United States

Related Subject Headings

  • Tetrahymena thermophila
  • Substrate Specificity
  • RNA, Messenger
  • RNA, Catalytic
  • Nucleic Acid Conformation
  • Molecular Sequence Data
  • Models, Chemical
  • Models, Biological
  • Kinetics
  • Introns