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Escherichia coli AlkB interacts with single-stranded DNA binding protein SSB by an intrinsically disordered region of SSB

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Abstract

Intrinsically disordered regions (IDRs) of proteins often regulate function through interactions with folded domains. Escherichia coli single-stranded DNA binding protein SSB binds and stabilizes single-stranded DNA (ssDNA). The N-terminal of SSB contains characteristic OB (oligonucleotide/oligosaccharide-binding) fold which binds ssDNA tightly but non-specifically. SSB also forms complexes with a large number proteins via the C-terminal interaction domain consisting mostly of acidic amino acid residues. The amino acid residues located between the OB-fold and C-terminal acidic domain are known to constitute an IDR and no functional significance has been attributed to this region. Although SSB is known to bind many DNA repair protein, it is not known whether it binds to DNA dealkylation repair protein AlkB. Here, we characterize AlkB SSB interaction and demonstrate that SSB binds to AlkB via the IDR. We have established that AlkB-SSB interaction by in vitro pull-down and yeast two-hybrid analysis. We mapped the site of contact to be the residues 152–169 of SSB. Unlike most of the SSB-binding proteins which utilize C-terminal acidic domain for interaction, IDR of SSB is necessary and sufficient for AlkB interaction.

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Funding

The work was funded by Extra-mural research Grant (EMR/2016/005135) funded by Science and Engineering Research Board (SERB), Government of India.

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  1. Richa Nigam and Monisha Mohan—equally contributed.

Authors and Affiliations

  1. Department of Biotechnology, Indian Institute of Technology Hyderabad, Kandi, Sangareddy, Telangana, 502285, India

    Richa Nigam, Monisha Mohan, Gururaj Shivange, Pranjal Kumar Dewangan & Roy Anindya

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  1. Richa Nigam
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  2. Monisha Mohan
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  3. Gururaj Shivange
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  4. Pranjal Kumar Dewangan
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Correspondence to Roy Anindya.

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Nigam, R., Mohan, M., Shivange, G. et al. Escherichia coli AlkB interacts with single-stranded DNA binding protein SSB by an intrinsically disordered region of SSB. Mol Biol Rep 45, 865–870 (2018). https://doi.org/10.1007/s11033-018-4232-6

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  • DOI: https://doi.org/10.1007/s11033-018-4232-6

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