Solution structure and DNA-binding properties of the winged helix domain of the meiotic recombination HOP2 protein.
The HOP2 protein is required for efficient double-strand break repair which ensures the proper synapsis of homologous chromosomes and normal meiotic progression. We previously showed that in vitro HOP2 shows two distinctive activities: when it is incorporated into a HOP2-MND1 heterodimer, it stimulates DMC1 and RAD51 recombination activities, and the purified HOP2 alone is proficient in promoting strand invasion. The structural and biochemical basis of HOP2 action in recombination are poorly understood; therefore, they are the focus of this work. Herein, we present the solution structure of the amino-terminal portion of mouse HOP2, which contains a typical winged helix DNA-binding domain. Together with NMR spectral changes in the presence of double-stranded DNA, protein docking on DNA, and mutation analysis to identify the amino acids involved in DNA coordination, our results on the three-dimensional structure of HOP2 provide key information on the fundamental structural and biochemical requirements directing the interaction of HOP2 with DNA. These results, in combination with mutational experiments showing the role of a coiled-coil structural feature involved in HOP2 self-association, allow us to explain important aspects of the function of HOP2 in recombination.
From the Department of Physics, Oklahoma State University, Stillwater, Oklahoma 74078.
Hem
H
Moktan
the Cell Cycle and Cancer Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104.
Michel F
MF
Guiraldelli
the Cell Cycle and Cancer Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104.
Craig A
CA
Eyster
the Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520.
Weixing
W
Zhao
the Cell Cycle and Cancer Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104.
Chih-Ying
CY
Lee
the Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, the Department of Biochemistry and Molecular Biology, Oklahoma University Health Sciences Center, Oklahoma City, Oklahoma 73104.
Timothy
T
Mather
the Genetics and Biochemistry Branch, NIDDK, National Institutes of Health, Bethesda, Maryland 20892, and.
R Daniel
RD
Camerini-Otero
the Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520.
Patrick
P
Sung
From the Department of Physics, Oklahoma State University, Stillwater, Oklahoma 74078.
Donghua H
DH
Zhou
the Cell Cycle and Cancer Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, the Department of Cell Biology, Oklahoma University Health Science Center, Oklahoma City, Oklahoma 73126 Roberto-Pezza@omrf.org.
Roberto J
RJ
Pezza
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D012996:Solutions / Q000737:chemistry
D000596:Amino Acids; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C501202:Hop2 protein, mouse; D012996:Solutions; D004247:DNA
DNA Binding; DNA Repair; DNA-binding Protein; Homologous Recombination; NMR; Protein Structure