Are the intrinsically disordered linkers involved in SSB binding to accessory proteins?

Min Kyung Shinn, Alexander G. Kozlov, Binh Nguyen, Wlodek M. Bujalowski, Timothy M. Lohman

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Escherichia coli single strand (ss) DNA binding (SSB) protein protects ssDNA intermediates and recruits at least 17 SSB interacting proteins (SIPs) during genome maintenance. The SSB C-termini contain a 9 residue acidic tip and a 56 residue intrinsically disordered linker (IDL). The acidic tip interacts with SIPs; however a recent proposal suggests that the IDL may also interact with SIPs. Here we examine the binding to four SIPs (RecO, PriC, PriA and χ subunit of DNA polymerase III) of three peptides containing the acidic tip and varying amounts of the IDL. Independent of IDL length, we find no differences in peptide binding to each individual SIP indicating that binding is due solely to the acidic tip. However, the tip shows specificity, with affinity decreasing in the order: RecO > PriA ∼ χ > PriC. Yet, RecO binding to the SSB tetramer and an SSB-ssDNA complex show significant thermodynamic differences compared to the peptides alone, suggesting that RecO interacts with another region of SSB, although not the IDL. SSB containing varying IDL deletions show different binding behavior, with the larger linker deletions inhibiting RecO binding, likely due to increased competition between the acidic tip interacting with DNA binding sites within SSB.

Original languageEnglish (US)
Pages (from-to)8581-8594
Number of pages14
JournalNucleic acids research
Volume47
Issue number16
DOIs
StatePublished - Sep 19 2019

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DNA
Proteins
DNA-Binding Proteins
Peptides
DNA Polymerase III
Thermodynamics
Protein Binding
Binding Sites
Maintenance
Genome
Escherichia coli

ASJC Scopus subject areas

  • Genetics

Cite this

Are the intrinsically disordered linkers involved in SSB binding to accessory proteins? / Shinn, Min Kyung; Kozlov, Alexander G.; Nguyen, Binh; Bujalowski, Wlodek M.; Lohman, Timothy M.

In: Nucleic acids research, Vol. 47, No. 16, 19.09.2019, p. 8581-8594.

Research output: Contribution to journalArticle

Shinn, Min Kyung ; Kozlov, Alexander G. ; Nguyen, Binh ; Bujalowski, Wlodek M. ; Lohman, Timothy M. / Are the intrinsically disordered linkers involved in SSB binding to accessory proteins?. In: Nucleic acids research. 2019 ; Vol. 47, No. 16. pp. 8581-8594.
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