The DNA Recognition Motif of GapR Has an Intrinsic DNA Binding Preference towards AT-rich DNA.

Qian Huang,Bin Xia,Shilong Fan,Bo Duan,Zhi Qu

doi:10.3390/molecules26195776

Qian Huang, Bin Xia + Show 3 more

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https://doi.org/10.3390/molecules26195776

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Abstract

The nucleoid-associated protein GapR found in Caulobacter crescentus is crucial for DNA replication, transcription, and cell division. Associated with overtwisted DNA in front of replication forks and the 3′ end of highly-expressed genes, GapR can stimulate gyrase and topo IV to relax (+) supercoils, thus facilitating the movement of the replication and transcription machines. GapR forms a dimer-of-dimers structure in solution that can exist in either an open or a closed conformation. It initially binds DNA through the open conformation and then undergoes structural rearrangement to form a closed tetramer, with DNA wrapped in the central channel. Here, we show that the DNA binding domain of GapR (residues 1–72, GapRΔC17) exists as a dimer in solution and adopts the same fold as the two dimer units in the full-length tetrameric protein. It binds DNA at the minor groove and reads the spatial distribution of DNA phosphate groups through a lysine/arginine network, with a preference towards AT-rich overtwisted DNA. These findings indicate that the dimer unit of GapR has an intrinsic DNA binding preference. Thus, at the initial binding step, the open tetramer of GapR with two relatively independent dimer units can be more efficiently recruited to overtwisted regions.

Highlights

GapR is a nucleoid-associated protein (NAP) that is highly conserved throughout the α-proteobacteria
The binding of GapR to overtwisted DNA sequences in front of the replication forks or downstream highly transcribed genes stimulates gyrase and topo IV to remove the (+) supercoils that arise during the unwinding of the DNA duplex, facilitating replication and transcription [3,5]
We determined the crystal structure of GapR∆C17 and illuminated its DNA recognition mechanism through NMR titration-based Haddock modeling. These findings indicate that the dimer unit of GapR has an intrinsic DNA binding preference towards AT-rich overtwisted DNA, which does not need the formation of a closed tetramer

Summary

Introduction

GapR is a nucleoid-associated protein (NAP) that is highly conserved throughout the α-proteobacteria. GapR shows a preference towards AT-rich DNA and overtwisted DNA, with a cellcycle-dependent and transcription-activity-related dynamic chromosome-binding pattern [1,3,4]. It is more enriched around the origin of replication at the swarmer cell stage and moves with the replication fork after chromosome replication initiates [2]. It is associated with highly expressed genes and operons [3]. The binding of GapR to overtwisted DNA sequences in front of the replication forks or downstream highly transcribed genes stimulates gyrase and topo IV to remove the (+) supercoils that arise during the unwinding of the DNA duplex, facilitating replication and transcription [3,5]

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