MeCP2 nuclear dynamics in live neurons results from low and high affinity chromatin interactions

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Abstract

Methyl-CpG-binding-Protein 2 (MeCP2) is an abundant nuclear protein highly enriched in neurons. Here we report live-cell single-molecule imaging studies of the kinetic features of mouse MeCP2 at high spatial-temporal resolution. MeCP2 displays dynamic features that are distinct from both highly mobile transcription factors and immobile histones. Stable binding of MeCP2 in living neurons requires its methyl-binding domain and is sensitive to DNA modification levels. Diffusion of unbound MeCP2 is strongly constrained by weak, transient interactions mediated primarily by its AT-hook domains, and varies with the level of chromatin compaction and cell type. These findings extend previous studies of the role of the MeCP2 MBD in high affinity DNA binding to living neurons, and identify a new role for its AT-hooks domains as critical determinants of its kinetic behavior. They suggest that limited nuclear diffusion of MeCP2 in live neurons contributes to its local impact on chromatin structure and gene expression.

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All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

Francesco M Piccolo

Laboratory of Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, United States

For correspondence
fpiccolo@rockefeller.edu

Competing interests
No competing interests declared.
Zhe Liu

Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-3592-3150
Peng Dong

Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States

Competing interests
No competing interests declared.
Ching-Lung Hsu

Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States

Competing interests
No competing interests declared.
Elitsa I Stoyanova

Laboratory of Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-6400-6119
Anjana Rao

Division of Signaling and Gene Expression, La Jolla Institute For Allergy and Immunology, La Jolla, United States

Competing interests
No competing interests declared.
Robert Tjian

Department of Molecular and Cell Biology, Li Ka Shing Center for Biomedical and Health Sciences, CIRM Center of Excellence, Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States

Competing interests
Robert Tjian, One of the three founding funders of eLife and a member of eLife's Board of Directors.

"This ORCID iD identifies the author of this article:" 0000-0003-0539-8217
Nathaniel Heintz

Laboratory of Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, United States

For correspondence
heintz@rockefeller.edu

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-8874-8704

Funding

Howard Hughes Medical Institute

Nathaniel Heintz

Howard Hughes Medical Institute

Zhe Liu

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (# 16944) of the Rockefeller University. The protocol was approved by the Committee on the Ethics of Animal Experiments of the Rockefeller University (OLAW assurance # #A3081-01).

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.