Structural insights into the recruitment of SMRT by the corepressor SHARP under phosphorylative regulation

Suzuka Mikami; Teppei Kanaba; Naoki Takizawa; Ayaho Kobayashi; Ryoko Maesaki; Toshinobu Fujiwara; Yutaka Ito; Masaki Mishima

doi:10.1016/j.str.2013.10.007

Structural insights into the recruitment of SMRT by the corepressor SHARP under phosphorylative regulation

Structure. 2014 Jan 7;22(1):35-46. doi: 10.1016/j.str.2013.10.007. Epub 2013 Nov 21.

Authors

Suzuka Mikami¹, Teppei Kanaba¹, Naoki Takizawa², Ayaho Kobayashi¹, Ryoko Maesaki³, Toshinobu Fujiwara², Yutaka Ito¹, Masaki Mishima⁴

Affiliations

¹ Graduate School of Science and Engineering, Tokyo Metropolitan University, 1-1 Minamiosawa Hachioji 192-0397, Japan.
² Institute of Microbial Chemistry, Tokyo 3-14-23 Kamiosaki, Shinagawa-ku 141-0021, Japan.
³ Graduate School of Science and Engineering, Tokyo Metropolitan University, 1-1 Minamiosawa Hachioji 192-0397, Japan; Graduate School of Bioscience, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
⁴ Graduate School of Science and Engineering, Tokyo Metropolitan University, 1-1 Minamiosawa Hachioji 192-0397, Japan. Electronic address: mishima-masaki@tmu.ac.jp.

PMID: 24268649
DOI: 10.1016/j.str.2013.10.007

Abstract

The transcriptional corepressors SMRT/NCoR, components of histone deacetylase complexes, interact with nuclear receptors and many other transcription factors. SMRT is a target for the ubiquitously expressed protein kinase CK2, which is known to phosphorylate a wide variety of substrates. Increasing evidence suggests that CK2 plays a regulatory role in many cellular events, particularly, in transcription. However, little is known about the precise mode of action involved. Here, we report the three-dimensional structure of a SMRT/HDAC1-associated repressor protein (SHARP) in complex with phosphorylated SMRT, as determined by solution NMR. Phosphorylation of the CK2 site on SMRT significantly increased affinity for SHARP. We also confirmed the significance of CK2 phosphorylation by reporter assay and propose a mechanism involving the process of phosphorylation acting as a molecular switch. Finally, we propose that the SPOC domain functions as a phosphorylation binding module.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Binding Sites
Casein Kinase II / chemistry*
Casein Kinase II / genetics
Casein Kinase II / metabolism
DNA-Binding Proteins
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression Regulation
Homeodomain Proteins / chemistry*
Homeodomain Proteins / genetics
Homeodomain Proteins / metabolism
Humans
Models, Molecular
Nuclear Magnetic Resonance, Biomolecular
Nuclear Proteins / chemistry*
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Nuclear Receptor Co-Repressor 2 / chemistry*
Nuclear Receptor Co-Repressor 2 / genetics
Nuclear Receptor Co-Repressor 2 / metabolism
Phosphorylation
Protein Binding
Protein Structure, Quaternary
Protein Structure, Tertiary
RNA-Binding Proteins
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Signal Transduction
Transcription, Genetic

Substances

DNA-Binding Proteins
Homeodomain Proteins
NCOR2 protein, human
Nuclear Proteins
Nuclear Receptor Co-Repressor 2
RNA-Binding Proteins
Recombinant Proteins
SPEN protein, human
Casein Kinase II