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Identification of novel regulatory domains in the yeast Snf2 protein
Author(s) -
Sen Payel,
Ghosh Sujana,
Pugh Frank,
Bartholomew Blaine
Publication year - 2010
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.24.1_supplement.830.3
Subject(s) - helicase , chromatin remodeling , atp hydrolysis , chromatin , yeast , atpase , protein subunit , saccharomyces cerevisiae , chemistry , microbiology and biotechnology , biochemistry , chromatin structure remodeling (rsc) complex , rna helicase a , swi/snf , transcription factor , aaa proteins , biology , dna , enzyme , rna , gene
The yeast SWI/SNF complex is a 1MDa multisubunit chromatin remodeler that uses ATP to alter chromatin structure for global transcription regulation. The ATPase‐helicase domain of the catalytic subunit Snf2 has been studied extensively but there has been little focus on regions outside this domain. This study identifies four domains outside the ATPase‐helicase domain that regulate catalytic activity, one of which also plays a role in targeting the complex for recruitment. Deletion of the first sixty amino acids (ΔN, 1–60) and the double AT‐hook (ΔAT, 1446–1530) decrease the rate of remodeling without much effect on ATP hydrolysis whereas deletion of spacer (1312–1444) and QLQ (239–527) show a severe defect in ATP hydrolysis. ΔN and ΔAT reduce remodeling efficiency by 13 and 29 fold respectively compared to WT, an extent that far exceeds their defect in ATP hydrolysis that is only 2.5 and 3.2 fold. Thus ΔN and ΔAT uncouple ATP hydrolysis from remodeling. The defects conferred by deletion of these domains are validated in vivo by growth assays and genome‐wide expression profiling. Biochemical characterization of complexes harboring point mutations in 9 amino acids conserved from yeast to humans in the spacer domain revealed moderate to severe remodeling defects. This study identifies novel, physiologically relevant domains and residues in Snf2 that regulate its targeting and catalytic function. NIH (GM48413)