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DNA Binding by a HEAT‐Repeat Subunit of Saccharomyces cerevisiae Condensin
Author(s) -
Sarkar Rupa,
Rybenkov Valentin V.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.912.6
Subject(s) - condensin , saccharomyces cerevisiae , biology , dna , protein subunit , microbiology and biotechnology , biochemistry , genetics , yeast , cohesin , chromatin , gene
Condensins play a crucial role in higher order chromosome organization in both prokaryotes and eukaryotes. In budding yeast Saccharomyces cerevisiae , condensin complex consists of five subunits: two evolutionary‐conserved structural maintenance of chromosome (SMC) subunits and three non‐SMC subunits. SMC subunits adopt a V‐shaped structure, in which two globular head domains are connected to a central hinge domain via two long coiled coils. Non‐SMC complex consists of a kleisin, which binds the head domains of SMC, and two HEAT‐repeat subunits Ycs4 and Ycg1. DNA binding and organization primarily resides in the SMC proteins. Non‐SMC subunits contribute to the activity of SMC and regulate the recruitment of holoenzyme to the chromosome. Unlike in bacteria, the non‐SMC complex of yeast S. cerevisiae binds DNA. In this study, we mapped the DNA binding site of non‐SMC complex using mass spectrometry based DNA footprinting. The DNA binding site is located on the C‐terminal domain of Ycs4 next to the HEAT‐repeat cluster. We verified the implicated amino acids using site‐directed mutagenesis and observed a reduction of DNA binding by Ycs4. Furthermore, we found that the mutations affect the cellular growth of S. cerevisiae and the activity of the holoenzyme condensin in vitro . These results show that DNA binding of Ycs4 is essential for the activity of condensin in vitro and in vivo . Support or Funding Information OCAST HR 14‐042