New clues in the allosteric activation of DNA cleavage by SgrAI

Structures of SgrAI bound to cleaved primary-site DNA and uncleaved secondary-site DNA

Elizabeth J. Little, Pete W. Dunten, Jurate Bitinaite, Nancy C Horton

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

SgrAI is a type II restriction endonuclease that cuts an unusually long recognition sequence and exhibits allosteric self-activation with expansion of DNA-sequence specificity. The three-dimensional crystal structures of SgrAI bound to cleaved primary-site DNA and Mg2+ and bound to secondary-site DNA with either Mg2+ or Ca2+ are presented. All three structures show a conformation of enzyme and DNA similar to the previously determined dimeric structure of SgrAI bound to uncleaved primary-site DNA and Ca2+ [Dunten et al. (2008), Nucleic Acids Res. 36, 5405-5416], with the exception of the cleaved bond and a slight shifting of the DNA in the SgrAI/cleaved primary-site DNA/Mg2+ structure. In addition, a new metal ion binding site is located in one of the two active sites in this structure, which is consistent with proposals for the existence of a metal-ion site near the 3′-O leaving group.

Original languageEnglish (US)
Pages (from-to)67-74
Number of pages8
JournalActa Crystallographica Section D: Biological Crystallography
Volume67
Issue number1
DOIs
StatePublished - Jan 2011

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DNA Cleavage
DNA
Type II Site Specific Deoxyribonucleases
Metals
Ions
Nucleic Acid Conformation
Nucleic Acids
Catalytic Domain
Binding Sites
Enzymes

Keywords

  • DNA cleavage
  • restriction endonucleases
  • SgrAI

ASJC Scopus subject areas

  • Structural Biology

Cite this

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title = "New clues in the allosteric activation of DNA cleavage by SgrAI: Structures of SgrAI bound to cleaved primary-site DNA and uncleaved secondary-site DNA",
abstract = "SgrAI is a type II restriction endonuclease that cuts an unusually long recognition sequence and exhibits allosteric self-activation with expansion of DNA-sequence specificity. The three-dimensional crystal structures of SgrAI bound to cleaved primary-site DNA and Mg2+ and bound to secondary-site DNA with either Mg2+ or Ca2+ are presented. All three structures show a conformation of enzyme and DNA similar to the previously determined dimeric structure of SgrAI bound to uncleaved primary-site DNA and Ca2+ [Dunten et al. (2008), Nucleic Acids Res. 36, 5405-5416], with the exception of the cleaved bond and a slight shifting of the DNA in the SgrAI/cleaved primary-site DNA/Mg2+ structure. In addition, a new metal ion binding site is located in one of the two active sites in this structure, which is consistent with proposals for the existence of a metal-ion site near the 3′-O leaving group.",
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T2 - Structures of SgrAI bound to cleaved primary-site DNA and uncleaved secondary-site DNA

AU - Little, Elizabeth J.

AU - Dunten, Pete W.

AU - Bitinaite, Jurate

AU - Horton, Nancy C

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AB - SgrAI is a type II restriction endonuclease that cuts an unusually long recognition sequence and exhibits allosteric self-activation with expansion of DNA-sequence specificity. The three-dimensional crystal structures of SgrAI bound to cleaved primary-site DNA and Mg2+ and bound to secondary-site DNA with either Mg2+ or Ca2+ are presented. All three structures show a conformation of enzyme and DNA similar to the previously determined dimeric structure of SgrAI bound to uncleaved primary-site DNA and Ca2+ [Dunten et al. (2008), Nucleic Acids Res. 36, 5405-5416], with the exception of the cleaved bond and a slight shifting of the DNA in the SgrAI/cleaved primary-site DNA/Mg2+ structure. In addition, a new metal ion binding site is located in one of the two active sites in this structure, which is consistent with proposals for the existence of a metal-ion site near the 3′-O leaving group.

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