Replication Protein A2 (RPA2) is a crucial subunit of the heterotrimeric Replication Protein A (RPA) complex. This complex plays a pivotal role in DNA metabolism, including DNA replication, repair, recombination, and telomere maintenance. RPA2 is also involved in coordinating the cellular response to DNA damage .
RPA2, also known as the 32 kDa subunit of the RPA complex, binds to single-stranded DNA (ssDNA) to form a nucleoprotein complex. This complex protects ssDNA from nucleases and prevents the formation of secondary structures that could interfere with DNA repair processes. The RPA complex has two modes of ssDNA binding: a low-affinity and a high-affinity mode, determined by the oligonucleotide/oligosaccharide-binding (OB) domains utilized .
Recombinant RPA2 is typically produced using bacterial expression systems. The gene encoding RPA2 is cloned into an expression vector, which is then introduced into a bacterial host such as Escherichia coli. The bacteria are cultured, and the recombinant protein is expressed and subsequently purified using affinity chromatography techniques .
RPA2 interacts with various proteins involved in genome maintenance. It contains a winged helix domain at its carboxy terminus, which facilitates interactions with other proteins. Post-translational modifications of the RPA complex, such as phosphorylation, play a significant role in coordinating different DNA damage response pathways .