Literature DB >> 2823642

A synthetic substrate for tRNA splicing.

V M Reyes1, J Abelson.   

Abstract

A novel method for the synthesis of precursor tRNA as substrate for in vitro splicing is reported. A construct consisting of the Saccharomyces cerevisiae pre-tRNAPhe gene under the control of a bacteriophage T7 promoter was assembled from a set of synthetic oligonucleotides and cloned into an M13 vector. By the use of T7 RNA polymerase, BstNI-runoff transcripts were produced. The resulting pre-tRNA was shown to possess mature termini and was accurately spliced by highly purified yeast tRNA-splicing endonuclease and ligase. Using this synthetic pre-tRNA, the kinetic parameters of the tRNA-splicing endonuclease were also determined. Use of this system provides several advantages for the study of tRNA-splicing mechanisms. Mutant tRNA precursors can be readily synthesized. It is also possible to synthesize large quantities of pre-tRNA for structural studies.

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Year:  1987        PMID： 2823642     DOI： 10.1016/0003-2697(87)90551-3

Source DB:  PubMed          Journal:  Anal Biochem        ISSN： 0003-2697            Impact factor:   3.365


  25 in total

1.  A primordial RNA modification enzyme: the case of tRNA (m1A) methyltransferase.

Authors:  Martine Roovers; Johan Wouters; Janusz M Bujnicki; Catherine Tricot; Victor Stalon; Henri Grosjean; Louis Droogmans
Journal:  Nucleic Acids Res       Date:  2004-01-22       Impact factor: 16.971

2.  The Cm56 tRNA modification in archaea is catalyzed either by a specific 2'-O-methylase, or a C/D sRNP.

Authors:  Marie-Hélène Renalier; Nicole Joseph; Christine Gaspin; Patricia Thebault; Annie Mougin
Journal:  RNA       Date:  2005-07       Impact factor: 4.942

3.  The use of a synthetic tRNA gene as a novel approach to study in vivo transcription and chromatin structure in yeast.

Authors:  R Krieg; R Stucka; S Clark; H Feldmann
Journal:  Nucleic Acids Res       Date:  1991-07-25       Impact factor: 16.971

4.  Characterization of elongating T7 and SP6 RNA polymerases and their response to a roadblock generated by a site-specific DNA binding protein.

Authors:  P A Pavco; D A Steege
Journal:  Nucleic Acids Res       Date:  1991-09-11       Impact factor: 16.971

5.  tRNA m7G methyltransferase Trm8p/Trm82p: evidence linking activity to a growth phenotype and implicating Trm82p in maintaining levels of active Trm8p.

Authors:  Andrei Alexandrov; Elizabeth J Grayhack; Eric M Phizicky
Journal:  RNA       Date:  2005-04-05       Impact factor: 4.942

6.  Cloning, in vitro transcription, and biological activity of Escherichia coli 23S ribosomal RNA.

Authors:  C J Weitzmann; P R Cunningham; J Ofengand
Journal:  Nucleic Acids Res       Date:  1990-06-25       Impact factor: 16.971

7.  Two proteins that form a complex are required for 7-methylguanosine modification of yeast tRNA.

Authors:  Andrei Alexandrov; Mark R Martzen; Eric M Phizicky
Journal:  RNA       Date:  2002-10       Impact factor: 4.942

8.  Cloning and characterization of tRNA (m1A58) methyltransferase (TrmI) from Thermus thermophilus HB27, a protein required for cell growth at extreme temperatures.

Authors:  Louis Droogmans; Martine Roovers; Janusz M Bujnicki; Catherine Tricot; Thomas Hartsch; Victor Stalon; Henri Grosjean
Journal:  Nucleic Acids Res       Date:  2003-04-15       Impact factor: 16.971

9.  Trans-acting RNA inhibits tRNA suppressor activity in vivo.

Authors:  Domenica Gandini Attardi; Glauco P Tocchini-Valentini
Journal:  RNA       Date:  2002-07       Impact factor: 4.942

10.  New archaeal methyltransferases forming 1-methyladenosine or 1-methyladenosine and 1-methylguanosine at position 9 of tRNA.

Authors:  Morgane Kempenaers; Martine Roovers; Yamina Oudjama; Karolina L Tkaczuk; Janusz M Bujnicki; Louis Droogmans
Journal:  Nucleic Acids Res       Date:  2010-06-04       Impact factor: 16.971
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