# Dynamic BTB domain filaments promote clustering of ZBTB proteins

Lucija Mance, Franck Coste, Sébastien Huet, Bertrand Castaing, Marcin Jozef

Józef Suskiewicz

► To cite this version:

Lucija Mance, Franck Coste, Sébastien Huet, Bertrand Castaing, Marcin Jozef Józef Suskiewicz. Dynamic BTB domain filaments promote clustering of ZBTB proteins. EMBO Meeting Ubiquitin and ubiquitin-like proteins in health and disease, Sep 2024, Cavtat & Dubrovnik, France. [hal-05216344](https://hal.science/hal-05216344v1)

HAL Id: hal-05216344

https://hal.science/hal-05216344v1

Submitted on 20 Aug 2025

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Distributed under a Creative Commons Attribution 4.0 International License
# Dynamic BTB domain filaments promote clustering of ZBTB proteins

Lucija Mance¹, Franck Coste¹, Sébastien Huet²,³, Bertrand Castaing¹, Marcin Józef Suskiewicz¹

¹ Centre de Biophysique Moléculaire (CBM), UPR 4301, CNRS, affiliated with Université d'Orléans, Orléans, France

² Institut de Génétique et Développement de Rennes (IGDR), UMR 6290, Université Rennes, CNRS, BIOSIT-UMS 3480, Rennes, France

³ Institut Universitaire de France, Paris, France

Email address: lucija.mance@cnrs-orleans.fr

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## INTRODUCTION

The ZBTB family (ZF and BTB domain-containing) comprises one of the largest transcription factor families in humans, with approximately 50 canonical members. Each ZBTB protein contains an N-terminal BTB domain that mediates dimerisation and a set of C-terminal zinc fingers responsible for DNA binding. Additionally, ZBTB proteins are prominent substrates for two posttranslational modifications - SUMOylation and ubiquitylation.

ZBTB proteins primarily function as transcriptional repressors, particularly in the context of differentiation and development. The two ZBTBs that we focus on in this study, **ZBTB8A (BOZF1)** and **ZBTB18 (RP58, ZNF238)**, also function as repressors. ZBTB8A is implicated in the development of prostate and breast cancer, while ZBTB18 is involved in neurodevelopmental disorders, intellectual disabilities, and cancer.

<figure><img src="image_10.png" /><figcaption>BTB domain dimer</figcaption></figure>

### Filamentation of BTB domain from ZBTB8A

<figure><img src="image_11.png" /><figcaption>The assembly of BTB filaments</figcaption></figure>

<figure><img src="image_12.png" /><figcaption>Size exclusion chromatography (Superdex 200)</figcaption></figure>

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### Structural insights into BTB interdimer interface

The BTB interdimer interface is symmetrical and it consists of two contacts.

<figure><img src="image_14.png" /><figcaption>Contact 1</figcaption></figure>

<figure><img src="image_15.png" /><figcaption>Contact 2</figcaption></figure>

<figure><img src="image_16.png" /></figure>

BTB filamentation dictates ZBTB subcellular localisation and mediates repression of a reporter gene

<figure><img src="image_17.png" /><figcaption>BTB filaments in solution</figcaption></figure>

Resolution: 3.1 Å
PDB: 8P2N

<figure><img src="image_18.png" /><figcaption>HsZBTB18</figcaption></figure>

<figure><img src="image_19.png" /><figcaption>Dual luciferase reporter assay</figcaption></figure>

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ZBTB18 binding site (BS10 sequence)

## CONCLUSIONS

* Our study reveals a previously unreported property of BTB domains of **ZBTB8A** and **ZBTB18** to form filaments

* The propensity for filamentation among certain human ZBTB transcription factors is required for the localisation of these proteins to nuclear foci in cells

* BTB domain-mediated polymerisation might potentially have functional significance in processes such as recognition of specific DNA motifs and gene repression.

<figure><img src="image_21.png" /></figure>

## PUBLICATION

Mance, L., Bigot, N., Sánchez, E. Z., Coste, F., Martín-González, N., Zentout, S., ... & Suskiewicz, M. J. (2024). Dynamic BTB-domain filaments promote clustering of ZBTB proteins. *Molecular Cell*, **84**(13), 2490-2510.

This result is part of a project that has received funding from the European Research Council (ERC) under the European Union's Horizon Europe programme, Starting Grant SUMOwriteNread, Grant agreement ID: 101078837.