Single-Molecule Force Spectroscopy Studies of APOBEC3A-Single-Stranded DNA Complexes

Biochemistry. 2016 Jun 7;55(22):3102-6. doi: 10.1021/acs.biochem.6b00214. Epub 2016 May 19.

Abstract

APOBEC3A (A3A) inhibits the replication of a range of viruses and transposons and might also play a role in carcinogenesis. It is a single-domain deaminase enzyme that interacts with single-stranded DNA (ssDNA) and converts cytidines to uridines within specific trinucleotide contexts. Although there is abundant information that describes the potential biological activities of A3A, the interplay between binding ssDNA and sequence-specific deaminase activity remains controversial. Using a single-molecule atomic force microscopy spectroscopy approach developed by Shlyakhtenko et al. [(2015) Sci. Rep. 5, 15648], we determine the stability of A3A in complex with different ssDNA sequences. We found that the strength of the complex is sequence-dependent, with more stable complexes formed with deaminase-specific sequences. A correlation between the deaminase activity of A3A and the complex strength was identified. The ssDNA binding properties of A3A and those for A3G are also compared and discussed.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cytidine Deaminase / chemistry*
  • Cytidine Deaminase / metabolism*
  • DNA, Single-Stranded / chemistry*
  • DNA, Single-Stranded / metabolism*
  • Deamination
  • Humans
  • Microscopy, Atomic Force / methods*
  • Protein Binding
  • Protein Conformation
  • Proteins / chemistry*
  • Proteins / metabolism*

Substances

  • DNA, Single-Stranded
  • Proteins
  • APOBEC3A protein, human
  • Cytidine Deaminase