The Intrinsically Disordered C-Terminal Tails of E. coli Single-Stranded DNA Binding Protein Regulate Cooperative Binding to Single-Stranded DNA

Козлов, А. Г.; Weiland, Elizabeth; Mittal, Anuradha; Waldman, Vince; Pappu, Rohit V.; Timothy, Lohman M.

doi:10.1016/j.bpj.2014.11.2132

Cited by 23 publications

(103 citation statements)

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“…Second, removal of these motifs should negatively impact cooperative binding to ssDNA. This is exactly what was observed by the Lohman group . Third, mutation of the PXXP motifs should negatively impact.…”

Section: Discussionsupporting

confidence: 79%

“…S1]. These are similar to those of the Lohman group, but differ in the deletion points and the length of the C‐terminal tip retained . For the proteins described herein, the subscript refers to the number of residues in each protein, counting the first methionine as position 1.…”

Section: Resultsmentioning

confidence: 95%

“…When the interactions take place between SSB tetramers, cooperative ssDNA binding occurs. When these key elements—the putative PPII‐helix, the repeats and the PXXP motifs are perturbed, SSB no longer functions correctly …”

Section: Discussionmentioning

confidence: 99%

“…The C‐terminal domain can be further subdivided into two additional regions: a sequence of ∼50 amino acids that has been called the intrinsically disordered linker (IDL) and the last 8–10 residues which are known as the acidic tip or C‐peptide and which is very well conserved among eubacterial proteins . While there is extensive knowledge of the N‐terminal domain, there are only a limited number of studies of the C‐terminus.…”

Section: Introductionmentioning

confidence: 99%

“…Further work both in vivo and in vitro using the ssbΔC8 gene and SSBΔC8 protein respectively, have shown that the last eight residues are critical, but not solely responsible, for target protein binding . One interpretation is that the acidic tip residues contact target proteins directly . An alternative proposal suggests that these last eight residues play a regulatory role and that target protein binding utilizes another region of SSB …”

Section: Introductionmentioning

confidence: 99%

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The IDL of E. coli SSB links ssDNA and protein binding by mediating protein–protein interactions

et al. 2017

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The E. coli single strand DNA binding protein (SSB) is essential to viability where it functions in two seemingly disparate roles: it binds to single stranded DNA (ssDNA) and to target proteins that comprise the SSB interactome. The link between these roles resides in a previously under-appreciated region of the protein known as the intrinsically disordered linker (IDL). We present a model wherein the IDL is responsible for mediating protein-protein interactions critical to each role. When interactions occur between SSB tetramers, cooperative binding to ssDNA results. When binding occurs between SSB and an interactome partner, storage or loading of that protein onto the DNA takes place. The properties of the IDL that facilitate these interactions include the presence of repeats, a putative polyproline type II helix and, PXXP motifs that may facilitate direct binding to the OB-fold in a manner similar to that observed for SH3 domain binding of PXXP ligands in eukaryotic systems.

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Section: Discussionsupporting

confidence: 79%

Section: Resultsmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The IDL of E. coli SSB links ssDNA and protein binding by mediating protein–protein interactions

et al. 2017

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SSB and the RecG DNA helicase: an intimate association to rescue a stalled replication fork

Bianco

Lyubchenko

2017

Protein Science

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In E. coli, the regression of stalled DNA replication forks is catalyzed by the DNA helicase RecG. One means of gaining access to the fork is by binding to the single strand binding protein or SSB. This interaction occurs via the wedge domain of RecG and the intrinsically disordered linker (IDL) of SSB, in a manner similar to that of SH3 domains binding to PXXP motif-containing ligands in eukaryotic cells. During loading, SSB remodels the wedge domain so that the helicase domains bind to the parental, duplex DNA, permitting the helicase to translocate using thermal energy. This translocation may be used to clear the fork of obstacles, prior to the initiation of fork regression.

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The intrinsically disordered linker of E. coliSSB is critical for the release from single‐stranded DNA

et al. 2017

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The Escherichia coli single stranded DNA binding protein (SSB) is crucial for DNA replication, recombination and repair. Within each process, it has two seemingly disparate roles: it stabilizes single-stranded DNA (ssDNA) intermediates generated during DNA processing and, forms complexes with a group of proteins known as the SSB-interactome. Key to both roles is the C-terminal, one-third of the protein, in particular the intrinsically disordered linker (IDL). Previously, they have shown using a series of linker deletion mutants that the IDL links both ssDNA and target protein binding by mediating interactions with the oligosaccharide/oligonucleotide binding fold in the target. In this study, they examine the role of the linker region in SSB function in a variety of DNA metabolic processes in vitro. Using the same linker mutants, the results show that in addition to association reactions (either DNA or protein), the IDL is critical for the release of SSB from DNA. This release can be under conditions of ssDNA competition or active displacement by a DNA helicase or recombinase. Consistent with their previous work these results indicate that SSB linker mutants are defective for SSB-SSB interactions, and when the IDL is removed a terminal SSB-DNA complex results. Formation of this complex inhibits downstream processing of DNA by helicases such as RecG or PriA as well as recombination, mediated by RecA. A model, based on the evidence herein, is presented to explain how the IDL acts in SSB function.

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The Intrinsically Disordered C-Terminal Tails of E. coli Single-Stranded DNA Binding Protein Regulate Cooperative Binding to Single-Stranded DNA

Cited by 23 publications

References 0 publications

The IDL of E. coli SSB links ssDNA and protein binding by mediating protein–protein interactions

The IDL of E. coli SSB links ssDNA and protein binding by mediating protein–protein interactions

SSB and the RecG DNA helicase: an intimate association to rescue a stalled replication fork

The intrinsically disordered linker of E. coliSSB is critical for the release from single‐stranded DNA

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