Solution structure of human Brg1 bromodomain and its specific binding to acetylated histone tails.
Human brahma-related gene 1 (Brg1) is a core protein in human SWI/SNF chromatin-remodeling complex which regulates gene expression. Brg1 contains a bromodomain that has been shown to anchor the entire complex to promoter nucleosomes by interacting with histones that are acetylated at specific lysine residues. The Brg1 bromodomain belongs to an important subclass of the bromodomain family for which no structural information is known. Here we report the solution structure of the Brg1 bromodomain determined by NMR. The Brg1 bromodomain conserves the left-handed, four-helix bundle topology found in other bromodomain structures. However, the alphaZ helix of Brg1 bromodomain is about 4 residues shorter relative to previously published bromodomain structures. Using NMR perturbation studies, we demonstrate the Brg1 bromodomain binds acetyllysine in the context of histone tails, with no comparable affinity for unacetylated peptides. The estimated dissociation constants (KD) for acetylated histone peptides H4-AcK8 and H4-AcK12 are 4.0 and 3.6 mM, respectively. In this study the dominant substrate was H3-AcK14 (KD approximately 1.2 mM). Mutagenesis analysis reveals several residues important for the binding specificity. Using molecular dynamics simulations, we present a model of the Brg1 bromodomain in complex with H3-AcK14 and discuss the potential interactions which provide the selectivity of the Brg1 bromodomain for histone H3-AcK14.
Shen
Weiqun
W
School of Life Sciences, and Hefei National Laboratory for Physical Sciences at Microscale, University of Science and Technology of China, Hefei, Anhui, 230026, People's Republic of China.
Xu
Chao
C
Huang
Wei
W
Zhang
Jiahai
J
Carlson
Justin E
JE
Tu
Xiaoming
X
Wu
Jihui
J
Shi
Yunyu
Y
D000107:Acetylation; D000595:Amino Acid Sequence; D002843:Chromatin / Q000378:metabolism; D004265:DNA Helicases / Q000737:chemistry* / Q000378:metabolism; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism
D002843:Chromatin; D006657:Histones; D009687:Nuclear Proteins; D014157:Transcription Factors; C084108:SMARCA4 protein, human; D004265:DNA Helicases