Molecular basis for SUMOylation-dependent regulation of DNA binding activity of heat shock factor 2.
Heat shock factor 2 (HSF2) is a member of a vertebrate transcription factor family for genes of heat shock proteins and is involved in the regulation of development and cellular differentiation. The DNA binding property of HSF2 is modulated by the post-translational modification of a specific lysine residue in its DNA binding domain by small ubiquitin-like modifier (SUMO), but the consequences of SUMOylation and its underlying molecular mechanism remain unclear. Here we show the inhibitory effect of SUMOylation on the interaction between HSF2 and DNA based on biochemical analysis using isolated recombinant HSF2. NMR study of the SUMOylated DNA binding domain of HSF2 indicates that the SUMO moiety is flexible with respect to the DNA binding domain and has neither a noncovalent interface with nor a structural effect on the domain. Combined with data from double electron-electron resonance and paramagnetic NMR relaxation enhancement experiments, these results suggest that SUMO attachment negatively modulates the formation of the protein-DNA complex through a randomly distributed steric interference.
Tateishi
Yukihiro
Y
Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan.
Ariyoshi
Mariko
M
Igarashi
Ryuji
R
Hara
Hideyuki
H
Mizuguchi
Kenji
K
Seto
Azusa
A
Nakai
Akira
A
Kokubo
Tetsuro
T
Tochio
Hidehito
H
Shirakawa
Masahiro
M
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D011485:Protein Binding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D025842:SUMO-1 Protein / Q000737:chemistry / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D006360:Heat-Shock Proteins; D025842:SUMO-1 Protein; D014157:Transcription Factors; C070389:HSF2 protein, human; D004247:DNA