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

O I Lavrik; D M Kolpashchikov; H P Nasheuer; K Weisshart; A Favre

doi:10.1016/s0014-5793(98)01544-0

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.

Authors

O I Lavrik¹, D M Kolpashchikov, H P Nasheuer, K Weisshart, A Favre

Affiliation

¹ Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Russian Academy of Sciences. lavrik@niboch.nsc.ru

PMID: 9883881
DOI: 10.1016/s0014-5793(98)01544-0

Abstract

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

Substances

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