Saccharomyces cerevisiae Red1 protein exhibits nonhomologous DNA end-joining activity and potentiates Hop1-promoted pairing of double-stranded DNA

被引:5
作者
Kshirsagar, Rucha [1 ]
Ghodke, Indrajeet [1 ]
Muniyappa, K. [1 ]
机构
[1] Indian Inst Sci, Dept Biochem, Bangalore 560012, Karnataka, India
关键词
YEAST HOP1 PROTEIN; SYNAPTONEMAL COMPLEX-FORMATION; MEIOTIC CHROMOSOME SYNAPSIS; SISTER-CHROMATID COHESION; AXIAL-ELEMENT PROTEIN; TELOMERIC DNA; BREAK REPAIR; MEK1-DEPENDENT PHOSPHORYLATION; INTERHOMOLOG RECOMBINATION; JUNCTION MIGRATION;
D O I
10.1074/jbc.M117.796425
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Elucidation of the function of synaptonemal complex (SC) in Saccharomyces cerevisiae has mainly focused on in vivo analysis of recombination-defective meiotic mutants. Consequently, significant gaps remain in the mechanistic understanding of the activities of various SC proteins and the functional relationships among them. S. cerevisiae Hop1 and Red1 are essential structural components of the SC axial/lateral elements. Previous studies have demonstrated that Hop1 is a structure-selective DNA-binding protein exhibiting high affinity for the Holliday junction and promoting DNA bridging, condensation, and pairing between double-stranded DNA molecules. However, the exact mode of action of Red1 remains unclear, although it is known to interact with Hop1 and to suppress the spore viability defects of hop1 mutant alleles. Here, we report the purification and functional characterization of the full-length Red1 protein. Our results revealed that Red1 forms a stable complex with Hop1 in vitro and provided quantitative insights into their physical interactions. Mechanistically, Red1 preferentially associated with the Holliday junction and 3-way junction rather than with single-or double-stranded DNA with overhangs. Although Hop1 and Red1 exhibited similar binding affinities toward several DNA substrates, the two proteins displayed some significant differences. Notably, Red1, by itself, lacked DNA-pairing ability; however, it potentiated Hop1-promoted intermolecular pairing between double-stranded DNA molecules. Moreover, Red1 exhibited nonhomologous DNA end-joining activity, thus revealing an unexpected role for Red1 in recombination-based DNA repair. Collectively, this study presents the first direct insights into Red1's mode of action and into the mechanism underlying its role in chromosome synapsis and recombination.
引用
收藏
页码:13853 / 13866
页数:14
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