Allosteric regulation of DNA cleavage and sequence-specificity through run-on oligomerization

Dmitry Lyumkis, Heather Talley, Andrew Stewart, Santosh Shah, Chad K. Park, Florence Tama, Clinton S. Potter, Bridget Carragher, Nancy C Horton

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

SgrAI is a sequence specific DNA endonuclease that functions through an unusual enzymatic mechanism that is allosterically activated 200- to 500-fold by effector DNA, with a concomitant expansion of its DNA sequence specificity. Using single-particle transmission electron microscopy to reconstruct distinct populations of SgrAI oligomers, we show that in the presence of allosteric, activating DNA, the enzyme forms regular, repeating helical structures characterized by the addition of DNA-binding dimeric SgrAI subunits in a run-on manner. We also present the structure of oligomeric SgrAI at 8.6 Å resolution, demonstrating the conformational state of SgrAI in its activated form. Activated and oligomeric SgrAI displays key protein-protein interactions near the helix axis between its N termini, as well as allosteric protein-DNA interactions that are required for enzymatic activation. The hybrid approach reveals an unusual mechanism of enzyme activation that explains SgrAI's oligomerization and allosteric behavior.

Original languageEnglish (US)
Pages (from-to)1848-1858
Number of pages11
JournalStructure
Volume21
Issue number10
DOIs
StatePublished - Oct 8 2013

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Allosteric Regulation
DNA Cleavage
DNA
Proteins
Enzyme Activation
Deoxyribonuclease I
Transmission Electron Microscopy
Enzymes
Population

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

Cite this

Allosteric regulation of DNA cleavage and sequence-specificity through run-on oligomerization. / Lyumkis, Dmitry; Talley, Heather; Stewart, Andrew; Shah, Santosh; Park, Chad K.; Tama, Florence; Potter, Clinton S.; Carragher, Bridget; Horton, Nancy C.

In: Structure, Vol. 21, No. 10, 08.10.2013, p. 1848-1858.

Research output: Contribution to journalArticle

Lyumkis, D, Talley, H, Stewart, A, Shah, S, Park, CK, Tama, F, Potter, CS, Carragher, B & Horton, NC 2013, 'Allosteric regulation of DNA cleavage and sequence-specificity through run-on oligomerization', Structure, vol. 21, no. 10, pp. 1848-1858. https://doi.org/10.1016/j.str.2013.08.012
Lyumkis D, Talley H, Stewart A, Shah S, Park CK, Tama F et al. Allosteric regulation of DNA cleavage and sequence-specificity through run-on oligomerization. Structure. 2013 Oct 8;21(10):1848-1858. https://doi.org/10.1016/j.str.2013.08.012
Lyumkis, Dmitry ; Talley, Heather ; Stewart, Andrew ; Shah, Santosh ; Park, Chad K. ; Tama, Florence ; Potter, Clinton S. ; Carragher, Bridget ; Horton, Nancy C. / Allosteric regulation of DNA cleavage and sequence-specificity through run-on oligomerization. In: Structure. 2013 ; Vol. 21, No. 10. pp. 1848-1858.
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