4.7 Article

Conformational dynamics promotes disordered regions from function-dispensable to essential in evolved site-specific DNA recombinases

Journal

COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL
Volume 20, Issue -, Pages 989-1001

Publisher

ELSEVIER
DOI: 10.1016/j.csbj.2022.01.010

Keywords

Intrinsically disordered protein regions; site-specific DNA recombinase; Evolution; Molecular dynamics; Structural disorder; Thermodynamic stability; AI-based folding

Funding

  1. German Research Council [DFG PI 600/4-1]
  2. European Union [ERC 742133, H2020 UPGRADE 825825]
  3. BMBF GO-Bio [031B0633]

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Protein intrinsically disordered regions (IDRs) play crucial roles in molecular recognition and regulation. By investigating the disordered N-terminal tail of Cre enzyme, the study provides mechanistic insights into the relationship between structural disorder and protein function. The findings offer new understanding of the functional implications of IDRs and their potential applications in biotechnology and genomic medicine.
Protein intrinsically disordered regions (IDRs) play pivotal roles in molecular recognition and regulatory processes through structural disorder-to-order transitions. To understand and exploit the distinctive functional implications of IDRs and to unravel the underlying molecular mechanisms, structural disorder-to-function relationships need to be deciphered. The DNA site-specific recombinase system Cre/loxP represents an attractive model to investigate functional molecular mechanisms of IDRs. Cre contains a functionally dispensable disordered N-terminal tail, which becomes indispensable in the evolved Tre/loxLTR recombinase system. The difficulty to experimentally obtain structural information about this tail has so far precluded any mechanistic study on its involvement in DNA recombination. Here, we use in vitro and in silico evolution data, conformational dynamics, AI-based folding simulations, thermodynamic stability calculations, mutagenesis and DNA recombination assays to investigate how evolution and the dynamic behavior of this IDR may determine distinct functional properties. Our studies suggest that partial conformational order in the N-terminal tail of Tre recombinase and its packing to a conserved hydrophobic surface on the protein provide thermodynamic stability. Based on our results, we propose a link between protein stability and function, offering new plausible atom-detailed mechanistic insights into disorder-function relationships. Our work highlights the potential of N-terminal tails to be exploited for regulation of the activity of Cre-like tyrosine-type SSRs, which merits future investigations and could be of relevance in future rational engineering for their use in biotechnology and genomic medicine. (c) 2022 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4.0/).

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