Alternative conformations of human replication protein A are detected by crosslinks with primers carrying a photoreactive group at the 3'-end

FEBS Lett. 1998 Dec 18;441(2):186-90. doi: 10.1016/s0014-5793(98)01544-0.


To analyze the influence of single-stranded template extension of DNA duplex on the conformation of human replication protein A (RPA) bound to DNA we have designed two template-primer systems differing by the size of the single-stranded template tail (9 and 19 nucleotides (nt)). Base-substituted photoreactive dUTP analogs were used as substrates for elongation of radiolabeled template-primer by DNA polymerase beta in the absence or in the presence of RPA. Following UV-crosslinking it was demonstrated that the pattern of RPA subunit labeling and consequently RPA arrangement near the 3'-end of the primer is strongly dependent upon the length of the template extension.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cross-Linking Reagents / chemistry
  • DNA Primers / chemistry*
  • DNA Replication
  • DNA-Binding Proteins / chemistry*
  • Humans
  • Photoaffinity Labels
  • Protein Conformation*
  • Replication Protein A
  • Templates, Genetic


  • Cross-Linking Reagents
  • DNA Primers
  • DNA-Binding Proteins
  • Photoaffinity Labels
  • RPA1 protein, human
  • Replication Protein A