HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ananvoranich, S. Articles by Perreault, J.-P. Search for Related Content PubMed PubMed Citation Articles by Ananvoranich, S. Articles by Perreault, J.-P. Social Bookmarking                      What's this?

J Biol Chem, Vol. 273, Issue 21, 13182-13188, May 22, 1998

Substrate Specificity of  Ribozyme Cleavage

From the Département de biochimie, Faculté de médecine, Université de Sherbrooke, Québec, J1H 5N4, Canada

The specificity of  ribozyme cleavage was investigated using a trans-acting antigenomic  ribozyme. Under single turnover conditions, the wild type ribozyme cleaved the 11-mer ribonucleotide substrate with a rate constant of 0.34 min¹, an apparent K_m of 17.9 nM and an apparent second-order rate constant of 1.89 × 10⁷ min¹ M¹. The substrate specificity of the  ribozyme was thoroughly investigated using a collection of substrates that varied in either the length or the nucleotide sequence of their P1 stems. We observed that not only is the base pairing of the substrate and the ribozyme important to cleavage activity, but also both the identity and the combination of the nucleotide sequence in the substrates are essential for cleavage activity. We show that the nucleotides in the middle of the P1 stem are essential for substrate binding and subsequent steps in the cleavage pathway. The introduction of any mismatches at these positions resulted in a complete lack of cleavage by the wild type ribozyme. Our findings suggest that factors more complex than simple base pairing interactions, such as tertiary structure interactions, could play an important role in the substrate specificity of  ribozyme cleavage.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				A. Nehdi, J. Perreault, J.-D. Beaudoin, and J.-P. Perreault A novel structural rearrangement of hepatitis delta virus antigenomic ribozyme Nucleic Acids Res., November 29, 2007; 35(20): 6820 - 6831. [Abstract] [Full Text] [PDF]

				A. Nehdi and J.-P. Perreault Unbiased in vitro selection reveals the unique character of the self-cleaving antigenomic HDV RNA sequence Nucleic Acids Res., January 23, 2006; 34(2): 584 - 592. [Abstract] [Full Text] [PDF]

				F. D'Anjou, L. J. Bergeron, N. B. Larbi, I. Fournier, M. Salzet, J.-P. Perreault, and R. Day Silencing of SPC2 Expression Using an Engineered {delta} Ribozyme in the Mouse {beta}TC-3 Endocrine Cell Line J. Biol. Chem., April 2, 2004; 279(14): 14232 - 14239. [Abstract] [Full Text] [PDF]

				P. Deschenes, J. Ouellet, J. Perreault, and J.-P. Perreault Formation of the P1.1 pseudoknot is critical for both the cleavage activity and substrate specificity of an antigenomic trans-acting hepatitis delta ribozyme Nucleic Acids Res., April 15, 2003; 31(8): 2087 - 2096. [Abstract] [Full Text] [PDF]

				L. J. Bergeron and J.-P. Perreault Development and comparison of procedures for the selection of delta ribozyme cleavage sites within the hepatitis B virus Nucleic Acids Res., November 1, 2002; 30(21): 4682 - 4691. [Abstract] [Full Text] [PDF]

				K. Fiola and J.-P. Perreault Kinetic and Binding Analysis of the Catalytic Involvement of Ribose Moieties of a trans-Acting delta Ribozyme J. Biol. Chem., July 12, 2002; 277(29): 26508 - 26516. [Abstract] [Full Text] [PDF]

				Y. Kato, T. Kuwabara, M. Warashina, H. Toda, and K. Taira Relationships between the Activities in Vitro and in Vivo of Various Kinds of Ribozyme and Their Intracellular Localization in Mammalian Cells J. Biol. Chem., April 27, 2001; 276(18): 15378 - 15385. [Abstract] [Full Text] [PDF]