DOI: 10.1101/486555Dec 4, 2018Paper

Recycling of single-stranded DNA-binding protein by the bacterial replisome.

BioRxiv : the Preprint Server for Biology
Lisanne M SpenkelinkAntoine M van Oijen

Abstract

Single-stranded DNA-binding proteins (SSBs) support DNA replication by protecting single-stranded DNA from nucleolytic attack, preventing intra-strand pairing events, and playing many other regulatory roles within the replisome. Recent developments in single-molecule approaches have led to a revised picture of the replisome that is much more complex in how it retains or recycles protein components. Here we visualise how an in vitro reconstituted E. coli replisome recruits SSB by relying on two different molecular mechanisms. Not only does it recruit new SSB molecules from solution to coat newly formed single-stranded DNA on the lagging strand, but it also internally recycles SSB from one Okazaki fragment to the next. We show that this internal transfer mechanism is balanced against recruitment from solution in a manner that is concentration dependent. By visualising SSB dynamics in live cells, we show that both internal transfer and external exchange mechanisms are physiologically relevant.

Related Concepts

DNA, Single-Stranded
DNA-Binding Proteins
Escherichia coli
Cerebrovascular Accident
Extrinsic
Approach
DNA Replication
Replisome
Small Molecule
probe gene fragment

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