Discovery and characterization of a thermostable bacteriophage RNA ligase homologous to T4 RNA ligase 1

Nucleic Acids Res. 2003 Dec 15;31(24):7247-54. doi: 10.1093/nar/gkg914.

Abstract

Thermophilic viruses represent a novel source of genetic material and enzymes with great potential for use in biotechnology. We have isolated a number of thermophilic viruses from geothermal areas in Iceland, and by combining high throughput genome sequencing and state of the art bioinformatics we have identified a number of genes with potential use in biotechnology. We have also demonstrated the existence of thermostable counterparts of previously known bacteriophage enzymes. Here we describe a thermostable RNA ligase 1 from the thermophilic bacteriophage RM378 that infects the thermophilic eubacterium Rhodothermus marinus. The RM378 RNA ligase 1 has a temperature optimum of 60-64 degrees C and it ligates both RNA and single-stranded DNA. Its thermostability and ability to work under conditions of high temperature where nucleic acid secondary structures are removed makes it an ideal enzyme for RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE), and other RNA and DNA ligation applications.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Amino Acid Sequence
  • Bacteriophages / enzymology*
  • Bacteriophages / genetics
  • Biotechnology
  • Cloning, Molecular
  • DNA, Single-Stranded / metabolism
  • Enzyme Stability
  • Hot Temperature
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • RNA / metabolism
  • RNA Ligase (ATP) / chemistry
  • RNA Ligase (ATP) / genetics
  • RNA Ligase (ATP) / isolation & purification
  • RNA Ligase (ATP) / metabolism*
  • Rhodothermus / virology*
  • Sequence Alignment
  • Substrate Specificity
  • Viral Proteins / metabolism*

Substances

  • DNA, Single-Stranded
  • Viral Proteins
  • RNA
  • Adenosine Triphosphate
  • RNA Ligase (ATP)
  • bacteriophage T4 RNA ligase 1

Associated data

  • RefSeq/NC_004735