Functions of Single-Strand DNA-Binding Proteins in DNA Replication, Recombination, and Repair

Abstract

Double-stranded (ds) DNA contains all of the necessary genetic information, although practical use of this information requires unwinding of the duplex DNA. DNA unwinding creates single-stranded (ss) DNA intermediates that serve as templates for myriad cellular functions. Exposure of ssDNA presents several problems to the cell. First, ssDNA is thermodynamically less stable than dsDNA, which leads to spontaneous formation of duplex secondary structures that impede genome maintenance processes. Second, relative to dsDNA, ssDNA is hypersensitive to chemical and nucleolytic attacks that can cause damage to the genome. Cells deal with these potential problems by encoding specialized ssDNA-binding proteins (SSBs) that bind to and stabilize ssDNA structures required for essential genomic processes. SSBs are essential proteins found in all domains of life. SSBs bind ssDNA with high affinity and in a sequence-independent manner and, in doing so, SSBs help to form the central nucleoprotein complex substrate for DNA replication, recombination, and repair processes. While SSBs are found in every organism, the proteins themselves share surprisingly little sequence similarity, subunit composition, and oligomerization states. All SSB proteins contain at least one DNA-binding oligonucleotide/oligosaccharide binding (OB) fold, which consists minimally of a five stranded beta-sheet arranged as a beta barrel capped by a single alpha helix. The OB fold is responsible for both ssDNA binding and oligomerization (for SSBs that operate as oligomers). The overall organization of OB folds varies between bacteria, eukaryotes, and archaea. As part of SSB/ssDNA cellular structures, SSBs play direct roles in the DNA replication, recombination, and repair. In many cases, SSBs have been found to form specific complexes with diverse genome maintenance proteins, often helping to recruit SSB/ssDNA-processing enzymes to the proper cellular sites of action. This clustering of genome maintenance factors can help to stimulate and coordinate the activities of individual enzymes and is also important for dislodging SSB from ssDNA. These features support a model in which DNA metabolic processes have evolved to work on ssDNA/SSB nucleoprotein filaments rather than on naked ssDNA. In this volume, methods are described to interrogate SSB-DNA and SSB-protein binding functions along with approaches that aim to understand the cellular functions of SSB. This introductory chapter offers a general overview of SSBs that focuses on their structures, DNA-binding mechanisms, and protein-binding partners.

Correlation Force Spectroscopy for Single Molecule Measurements

“…ssDNA is an essential intermediate in many DNA metabolic processes such as replication, recombination, repair, and transcription, and it is specifically recognized by many DNA-binding proteins, and thus its behavior is of growing interest to the scientific community [124][125][126][127][128][129][130][131][132][133].…”

Replication protein A and more: single-stranded DNA-binding proteins in eukaryotic cells

“…Although they have a wide range of sequences and differ markedly in their subunit composition and oligomerization states, they all bind to ssDNA with high affinity and in a sequence-independent manner [1,2]. In this way, SSBs form nucleoprotein complexes with ssDNA, which serve as substrates for DNA replication, recombination, and repair processes.…”

“…As a result, single-stranded DNA (ssDNA) is probably one of the most abundant and important DNA intermediate structures in cells [1]. However, ssDNA is less stable and very easily forms spontaneous duplex DNA or is attacked by chemical and nucleolytic reagents.…”

“…Although genetic information is carried by double-stranded DNA (dsDNA), the practical use of that information requires the unwinding of duplex DNA [1,2]. As a result, single-stranded DNA (ssDNA) is probably one of the most abundant and important DNA intermediate structures in cells [1].…”

“…However, ssDNA is less stable and very easily forms spontaneous duplex DNA or is attacked by chemical and nucleolytic reagents. To deal with these risks, cells have evolved a group of protective ssDNA-binding proteins (SSBs) that bind ssDNA with high affinity and specificity [1,2].…”