Characterization of reverse transcriptase activity of the L1Tc retroelement from Trypanosoma cruzi

J L García-Pérez, C I González, M C Thomas, M Olivares, M C López

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

The recombinant protein RTL1Tc, encoded by the non-LTR (long terminal repeat) retrotransposon L1Tc from Trypanosoma cruzi, has been shown to have reverse transcriptase (RT) activity using poly(rA)/oligo(dT) and poly(rC)/oligo(dG) homopolymers as template/primers. The optimal RT activity was detected at a concentration of 5 mM Mg2+, pH 8 and between 28 and 37% degrees C. Site-directed mutagenesis in the RT catalytic site proved that substitution of aspartic acid 313 for isoleucine (RT D313IL1Tc) practically abolishes the RT activity of the RTL1Tc protein. RT-polymerase chain reaction assays revealed that the RTL1Tc protein has the ability to use both homologous and heterologous RNA templates. Also, it is shown that the RTL1Tc protein is capable of synthesizing complementary DNA molecules by consecutive switching of the oligo molecule, which the protein uses as a template. This template switching may be involved in the retroelement integration process.

Original languageEnglish
Pages (from-to)2692-701
Number of pages10
JournalCellular and Molecular Life Sciences
Volume60
Issue number12
DOIs
Publication statusPublished - Dec 2003

Keywords / Materials (for Non-textual outputs)

  • Animals
  • Base Sequence
  • Binding Sites
  • Cloning, Molecular
  • DNA, Complementary
  • Molecular Sequence Data
  • RNA-Directed DNA Polymerase
  • Retroelements
  • Trypanosoma cruzi

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