title: Functional attributes of the Saccharomyces cerevisiae meiotic recombinase Dmc1.
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 ∼106Å. The ScDmc1 purification procedure and companion molecular analyses should facilitate future studies on this recombinase.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06520, USA.
authors.forename: Valeria
authors.initials: V
authors.lastname: Busygina
authors.forename: William A
authors.initials: WA
authors.lastname: Gaines
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Xu
authors.forename: Youngho
authors.initials: Y
authors.lastname: Kwon
authors.forename: Gareth J
authors.initials: GJ
authors.lastname: Williams
authors.forename: Sheng-Wei
authors.initials: SW
authors.lastname: Lin
authors.forename: Hao-Yen
authors.initials: HY
authors.lastname: Chang
authors.forename: Peter
authors.initials: P
authors.lastname: Chi
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Wang
authors.forename: Patrick
authors.initials: P
authors.lastname: Sung
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D002118:Calcium / Q000737:chemistry; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D002850:Chromatography, Gel; D004265:DNA Helicases / Q000737:chemistry; D045643:DNA Repair Enzymes / Q000737:chemistry; D026942:DNA Topoisomerases / Q000737:chemistry; D004271:DNA, Fungal / Q000737:chemistry / Q000648:ultrastructure; D004277:DNA, Single-Stranded / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D059765:Homologous Recombination; D006801:Humans; D006868:Hydrolysis; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*
chemical_list: D018797:Cell Cycle Proteins; C513033:DMC1 protein, S cerevisiae; D004271:DNA, Fungal; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D000255:Adenosine Triphosphate; C075468:RAD54 protein, S cerevisiae; D004265:DNA Helicases; D026942:DNA Topoisomerases; C541336:RDH54 protein, S cerevisiae; D045643:DNA Repair Enzymes; D002118:Calcium
keywords: DSB; Dmc1 recombinase; HR; Homologous recombination; Meiosis; N-terminal domain; NTD; double-strand break; homologous recombination