The Essential, Ubiquitous Single-Stranded DNA-Binding Proteins

Oliveira, Marcos T.; Ciesielski, Grzegorz L.

doi:10.1007/978-1-0716-1290-3_1

Cited by 6 publications

(10 citation statements)

References 173 publications

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“…In this study, we aimed to evaluate the physical interaction between mtSSB and Pol γ, and demonstrated for the first time that mtSSB interacts physically with Pol γα. The fact that this interaction occurs in the absence of a C-terminal domain, which in cases of both the homologous SSB protein of E. coli or non-homologous SSBs of phages T7 and T4, is necessary to bind cognate DNA polymerases (Oliveira and Ciesielski, 2021), makes our finding unique. Notably, the interaction occurs only in the absence of the accessory subunit Pol γβ.…”

Section: Discussionmentioning

confidence: 96%

Physical and Functional Interaction of Mitochondrial Single-Stranded DNA-Binding Protein and the Catalytic Subunit of DNA Polymerase Gamma

et al. 2021

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The maintenance of the mitochondrial genome depends on a suite of nucleus-encoded proteins, among which the catalytic subunit of the mitochondrial replicative DNA polymerase, Pol γα, plays a pivotal role. Mutations in the Pol γα-encoding gene, POLG, are a major cause of human mitochondrial disorders. Here we present a study of direct and functional interactions of Pol γα with the mitochondrial single-stranded DNA-binding protein (mtSSB). mtSSB coordinates the activity of the enzymes at the DNA replication fork. However, the mechanism of this functional relationship is elusive, and no direct interactions between the replicative factors have been identified to date. This contrasts strikingly with the extensive interactomes of SSB proteins identified in other homologous replication systems. Here we show for the first time that mtSSB binds Pol γα directly, in a DNA-independent manner. This interaction is strengthened in the absence of the loop 2.3 structure in mtSSB, and is abolished upon preincubation with Pol γβ. Together, our findings suggest that the interaction between mtSSB and polymerase gamma holoenzyme (Pol γ) involves a balance between attractive and repulsive affinities, which have distinct effects on DNA synthesis and exonucleolysis.

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Section: Discussionmentioning

confidence: 96%

Physical and Functional Interaction of Mitochondrial Single-Stranded DNA-Binding Protein and the Catalytic Subunit of DNA Polymerase Gamma

et al. 2021

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“…S4 & Table 1). Most characterized SSBs have a high affinity and specificity for ssDNA 27 . As an example, RPA binds mixed ssDNA with affinities of 10-40 nM albeit displaying a preference to pyrimidines, and with KD values to ssDNA up to 3 orders of magnitude lower than to dsDNA [51][52][53] .…”

Section: Discussionmentioning

confidence: 99%

PrgE: an OB-fold protein from plasmid pCF10 with striking differences to prototypical bacterial SSBs

Breidenstein,

Lamy,

Bader

et al. 2024

Preprint

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A major pathway for horizontal gene transfer is the transmission of DNA from donor to recipient cells via plasmid-encoded Type 4 Secretion Systems (T4SS). Many conjugative plasmids encode for a single-stranded DNA-binding protein (SSB) together with their T4SS. Some of these SSBs have been suggested to aid in establishing the plasmid in the recipient cell, but for many their function remains unclear. Here, we characterize PrgE, a proposed SSB from Enterococcus faecalis plasmid pCF10. We show that PrgE is not essential for conjugation. Structurally, it has the characteristic OB-fold of SSBs, but it has very uncharacteristic DNA-binding properties. Our DNA-bound structure shows that PrgE binds ssDNA like beads on a string, and this plasticity of PrgEs oligomerization is further confirmed by in vitro studies. Unlike other SSBs, PrgE binds both double- and single-stranded DNA equally well. This shows that PrgE has a quaternary assembly and DNA-binding properties that are very different from the prototypical bacterial SSB, but also different from the eukaryotic SSBs.

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“…Surprisingly, we found that PrgE bound dsDNA equally well as ssDNA (Figs 7 and S4 and Table 1). Most characterized SSBs have a high affinity and specificity for ssDNA (Oliveira & Ciesielski, 2021). As an example, RPA binds mixed ssDNA with affinities of 10-40 nM albeit displaying a preference for pyrimidines, and with K D values to ssDNA up to three orders of magnitude lower than to dsDNA (Brill & Stillman, 1989;Wold et al, 1989;Kim et al, 1992).…”

Section: Discussionmentioning

confidence: 99%

“…When it comes to archaea, some phyla have SSBs that resemble bacterial SSBs, whereas others are more similar to eukaryotic RPA (Taib et al, 2021). There are not only viruses that rely exclusively on host SSBs, but also those that encode their own proteins, with a large diversity of characteristics, some of which act as monomers (Shokri et al, 2009;Oliveira & Ciesielski, 2021). However, there is also variation within the kingdoms, as many bacterial and eukaryotic species have more than one type of OB-fold protein, which can vary significantly from their respective prototypes (Richard et al, 2008;Flynn & Zou, 2010;Yadav et al, 2012;Oliveira & Ciesielski, 2021).…”

Section: Introductionmentioning

confidence: 99%

PrgE: an OB-fold protein from plasmid pCF10 with striking differences to prototypical bacterial SSBs

Breidenstein,

Lamy,

Bader

et al. 2024

Life Sci. Alliance

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A major pathway for horizontal gene transfer is the transmission of DNA from donor to recipient cells via plasmid-encoded type IV secretion systems (T4SSs). Many conjugative plasmids encode for a single-stranded DNA-binding protein (SSB) together with their T4SS. Some of these SSBs have been suggested to aid in establishing the plasmid in the recipient cell, but for many, their function remains unclear. Here, we characterize PrgE, a proposed SSB from theEnterococcus faecalisplasmid pCF10. We show that PrgE is not essential for conjugation. Structurally, it has the characteristic OB-fold of SSBs, but it has very unusual DNA-binding properties. Our DNA-bound structure shows that PrgE binds ssDNA like beads on a string supported by its N-terminal tail. In vitro studies highlight the plasticity of PrgE oligomerization and confirm the importance of the N-terminus. Unlike other SSBs, PrgE binds both double- and single-stranded DNA equally well. This shows that PrgE has a quaternary assembly and DNA-binding properties that are very different from the prototypical bacterial SSB, but also different from eukaryotic SSBs.

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The Essential, Ubiquitous Single-Stranded DNA-Binding Proteins

Cited by 6 publications

References 173 publications

Physical and Functional Interaction of Mitochondrial Single-Stranded DNA-Binding Protein and the Catalytic Subunit of DNA Polymerase Gamma

Physical and Functional Interaction of Mitochondrial Single-Stranded DNA-Binding Protein and the Catalytic Subunit of DNA Polymerase Gamma

PrgE: an OB-fold protein from plasmid pCF10 with striking differences to prototypical bacterial SSBs

PrgE: an OB-fold protein from plasmid pCF10 with striking differences to prototypical bacterial SSBs

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