RPA-1 from Leishmania amazonensis (LaRPA-1) structurally differs from other eukaryote RPA-1 and interacts with telomeric DNA via its N-terminal OB-fold domain

FEBS Lett. 2014 Dec 20;588(24):4740-8. doi: 10.1016/j.febslet.2014.11.005. Epub 2014 Nov 15.

Abstract

Replication protein A-1 (RPA-1) is a single-stranded DNA-binding protein involved in DNA metabolism. We previously demonstrated the interaction between LaRPA-1 and telomeric DNA. Here, we expressed and purified truncated mutants of LaRPA-1 and used circular dichroism measurements and molecular dynamics simulations to demonstrate that the tertiary structure of LaRPA-1 differs from human and yeast RPA-1. LaRPA-1 interacts with telomeric ssDNA via its N-terminal OB-fold domain, whereas RPA from higher eukaryotes show different binding modes to ssDNA. Our results show that LaRPA-1 is evolutionary distinct from other RPA-1 proteins and can potentially be used for targeting trypanosomatid telomeres.

Keywords: LaRPA-1; Leishmania amazonensis; OB-fold domain; Replication factor 1; Replication protein A subunit 70kDa; Telomeres.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Computer Simulation
  • DNA, Single-Stranded / metabolism*
  • Humans
  • Leishmania*
  • Models, Molecular
  • Molecular Sequence Data
  • Oligonucleotides / metabolism
  • Oligosaccharides / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • Protozoan Proteins / chemistry*
  • Protozoan Proteins / metabolism*
  • Replication Protein A / chemistry*
  • Replication Protein A / metabolism*
  • Species Specificity
  • Telomere / genetics*

Substances

  • DNA, Single-Stranded
  • Oligonucleotides
  • Oligosaccharides
  • Protozoan Proteins
  • Replication Protein A