SSB binds to the RecG and PriA helicases in vivo in the absence of DNA

Genes to Cells : Devoted to Molecular & Cellular Mechanisms
Cong YuPiero R Bianco

Abstract

The E. coli single-stranded DNA-binding protein (SSB) binds to the fork DNA helicases RecG and PriA in vitro. Typically for binding to occur, 1.3 m ammonium sulfate must be present, bringing into question the validity of these results as these are nonphysiological conditions. To determine whether SSB can bind to these helicases, we examined binding in vivo. First, using fluorescence microscopy, we show that SSB localizes PriA and RecG to the vicinity of the inner membrane in the absence of DNA damage. Localization requires that SSB be in excess over the DNA helicases and the SSB C-terminus and both PriA and RecG be present. Second, using the purification of tagged complexes, our results show that SSB binds to PriA and RecG in vivo, in the absence of DNA. We propose that this may be the 'storage form' of RecG and PriA. We further propose that when forks stall, RecG and PriA are targeted to the fork by SSB, which, by virtue of its high affinity for single-stranded DNA, allows these helicases to outcompete other proteins. This ensures their actions in the early stages of the rescue of stalled replication forks.

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Citations

Aug 8, 2014·PLoS Genetics·Julia S P Mawer, David R F Leach
Oct 14, 2016·PloS One·Getachew Tesfaye BeyeneTone Tønjum
Nov 14, 2016·Progress in Biophysics and Molecular Biology·Piero R Bianco
Jan 13, 2017·Protein Science : a Publication of the Protein Society·Hui Yin TanPiero R Bianco
Jan 13, 2017·Protein Science : a Publication of the Protein Society·Piero R Bianco, Yuri L Lyubchenko
Dec 5, 2017·Nucleic Acids Research·Tricia A WindgassenJames L Keck
Jul 23, 2019·Nucleic Acids Research·Min Kyung ShinnTimothy M Lohman
Mar 21, 2020·Protein Science : a Publication of the Protein Society·Wenfei DingPiero R Bianco
Oct 23, 2019·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Tianyu ZhaoPiero R Bianco
Jan 28, 2017·Protein Science : a Publication of the Protein Society·Piero R BiancoUmesh Varshney
Mar 27, 2020·The Journal of Biological Chemistry·Yaqing WangYuri L Lyubchenko
Oct 7, 2020·Proceedings of the National Academy of Sciences of the United States of America·Gábor M HaramiMihály Kovács
Jul 28, 2019·Biophysical Journal·Bilyana N KolevaPenny J Beuning
Jan 16, 2021·Nucleic Acids Research·Min Kyung ShinnTimothy M Lohman
Mar 22, 2021·Nucleic Acids Research·Piero R Bianco, Yue Lu
Apr 22, 2021·The Journal of Physical Chemistry. B·Zhiqiang SunYuri L Lyubchenko
Jun 6, 2021·Protein Science : a Publication of the Protein Society·Piero R Bianco
Jul 9, 2021·ACS Omega·Hui Yin Tan, Piero R Bianco
Feb 13, 2018·Biochemistry·Zhiqiang SunYuri L Lyubchenko

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