SSB binds to the RecG and PriA helicases in vivo in the absence of DNA. (13th January 2016)

• Record Type: Journal Article
• Authors: Yu, Cong; Tan, Hui Yin; Choi, Meerim; Stanenas, Adam J.; Byrd, Alicia K.; D. Raney, Kevin; Cohan, Christopher S.; Bianco, Piero R.
• Is Part Of: Genes to cells. Volume 21:Number 2(2016)
• Page Start: 163
• Page End: 184
• Publication Date: 2016-01-13
• Subjects: Cytogenetics -- Periodicals; Cells -- Mechanical properties -- Periodicals; Molecular genetics -- Periodicals; Genes -- Periodicals; Molecular biology -- Periodicals; Cytology -- Periodicals; Biomechanics -- Periodicals; 571.6
• DOI: 10.1111/gtc.12334
• Languages: English
• ISSNs: 1356-9597

Abstract

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. Abstract : RecG and PriA bind to SSB in vivo in the absence of DNA. In this storage form, the helicases are found in the vicinity of the inner membrane. When needed, they are transferred to a stalled replication fork.