Functional attributes of the Saccharomyces cerevisiae meiotic recombinase Dmc1

Valeria Busygina, William A. Gaines, Yuanyuan Xu, Youngho Kwon, Gareth J. Williams, Sheng Wei Lin, Hao Yen Chang, Peter Chi, Hong Wei Wang, Patrick Sung

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

The role of Dmc1 as a meiosis-specific general recombinase was first demonstrated in Saccharomyces cerevisiae. Progress in understanding the biochemical mechanism of ScDmc1 has been hampered by its tendency to form inactive aggregates. We have found that the inclusion of ATP during protein purification prevents Dmc1 aggregation. ScDmc1 so prepared is capable of forming D-loops and responsive to its accessory factors Rad54 and Rdh54. Negative staining electron microscopy and iterative helical real-space reconstruction revealed that the ScDmc1-ssDNA nucleoprotein filament harbors 6.5 protomers per turn with a pitch of ~106. Å. The ScDmc1 purification procedure and companion molecular analyses should facilitate future studies on this recombinase.

Original languageEnglish (US)
Pages (from-to)707-712
Number of pages6
JournalDNA Repair
Volume12
Issue number9
DOIs
StatePublished - Sep 2013
Externally publishedYes

Keywords

  • Dmc1 recombinase
  • Homologous recombination
  • Meiosis

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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