Cryo-EM structure of CtBP2 confirms tetrameric architecture

Anne M Jecrois; M Michael Dcona; Xiaoyan Deng; Dipankar Bandyopadhyay; Steven R Grossman; Celia A Schiffer; William E Royer Jr

doi:10.1016/j.str.2020.11.008

Cryo-EM structure of CtBP2 confirms tetrameric architecture

Structure. 2021 Apr 1;29(4):310-319.e5. doi: 10.1016/j.str.2020.11.008. Epub 2020 Dec 1.

Authors

Anne M Jecrois¹, M Michael Dcona², Xiaoyan Deng³, Dipankar Bandyopadhyay⁴, Steven R Grossman⁵, Celia A Schiffer¹, William E Royer Jr⁶

Affiliations

¹ Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
² Department of Internal Medicine, Virginia Commonwealth University, Richmond, VA 23298, USA.
³ Department of Biostatistics, Virginia Commonwealth University, Richmond, VA 23298, USA.
⁴ Department of Biostatistics, Virginia Commonwealth University, Richmond, VA 23298, USA; Massey Cancer Center, Virginia Commonwealth University, Richmond, VA 23298, USA.
⁵ Department of Internal Medicine, Virginia Commonwealth University, Richmond, VA 23298, USA; Massey Cancer Center, Virginia Commonwealth University, Richmond, VA 23298, USA.
⁶ Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA. Electronic address: william.royer@umassmed.edu.

Abstract

C-terminal binding proteins 1 and 2 (CtBP1 and CtBP2) are transcriptional regulators that activate or repress many genes involved in cellular development, apoptosis, and metastasis. NADH-dependent CtBP activation has been implicated in multiple types of cancer and poor patient prognosis. Central to understanding activation of CtBP in oncogenesis is uncovering how NADH triggers protein assembly, what level of assembly occurs, and if oncogenic activity depends upon such assembly. Here, we present the cryoelectron microscopic structures of two different constructs of CtBP2 corroborating that the native state of CtBP2 in the presence of NADH is tetrameric. The physiological relevance of the observed tetramer was demonstrated in cell culture, showing that CtBP tetramer-destabilizing mutants are defective for cell migration, transcriptional repression of E-cadherin, and activation of TIAM1. Together with our cryoelectron microscopy studies, these results highlight the tetramer as the functional oligomeric form of CtBP2.

Keywords: C-terminal binding-protein CtBP; TIAM1; cancer; metastasis; tetrameric assembly; transcription regulation.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Alcohol Oxidoreductases / chemistry*
Alcohol Oxidoreductases / genetics
Alcohol Oxidoreductases / metabolism
Cadherins / metabolism
Catalytic Domain
Cell Movement
Co-Repressor Proteins / chemistry*
Co-Repressor Proteins / genetics
Co-Repressor Proteins / metabolism
Cryoelectron Microscopy
DNA-Binding Proteins / chemistry*
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Gene Expression Regulation, Neoplastic
HCT116 Cells
Humans
Mutation
NADP / metabolism
Protein Binding
Protein Multimerization*
T-Lymphoma Invasion and Metastasis-inducing Protein 1 / metabolism

Substances

Cadherins
Co-Repressor Proteins
DNA-Binding Proteins
T-Lymphoma Invasion and Metastasis-inducing Protein 1
TIAM1 protein, human
NADP
Alcohol Oxidoreductases
CTBP2 protein, human
C-terminal binding protein

Abstract

Publication types

MeSH terms

Substances

Grants and funding