Integrated requirement of non-specific and sequence-specific DNA binding in Myc-driven transcription.


Journal

The EMBO journal
ISSN: 1460-2075
Titre abrégé: EMBO J
Pays: England
ID NLM: 8208664

Informations de publication

Date de publication:
17 05 2021
Historique:
revised: 15 02 2021
received: 01 05 2020
accepted: 24 02 2021
pubmed: 2 4 2021
medline: 16 11 2021
entrez: 1 4 2021
Statut: ppublish

Résumé

Eukaryotic transcription factors recognize specific DNA sequence motifs, but are also endowed with generic, non-specific DNA-binding activity. How these binding modes are integrated to determine select transcriptional outputs remains unresolved. We addressed this question by site-directed mutagenesis of the Myc transcription factor. Impairment of non-specific DNA backbone contacts caused pervasive loss of genome interactions and gene regulation, associated with increased intra-nuclear mobility of the Myc protein in murine cells. In contrast, a mutant lacking base-specific contacts retained DNA-binding and mobility profiles comparable to those of the wild-type protein, but failed to recognize its consensus binding motif (E-box) and could not activate Myc-target genes. Incidentally, this mutant gained weak affinity for an alternative motif, driving aberrant activation of different genes. Altogether, our data show that non-specific DNA binding is required to engage onto genomic regulatory regions; sequence recognition in turn contributes to transcriptional activation, acting at distinct levels: stabilization and positioning of Myc onto DNA, and-unexpectedly-promotion of its transcriptional activity. Hence, seemingly pervasive genome interaction profiles, as detected by ChIP-seq, actually encompass diverse DNA-binding modalities, driving defined, sequence-dependent transcriptional responses.

Identifiants

pubmed: 33792944
doi: 10.15252/embj.2020105464
pmc: PMC8126922
doi:

Substances chimiques

Proto-Oncogene Proteins c-myc 0
Transcription Factors 0
DNA 9007-49-2

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Pagination

e105464

Informations de copyright

© 2021 The Authors.

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Auteurs

Paola Pellanda (P)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.
Center for Genomic Science of IIT@SEMM, Fondazione Istituto Italiano di Tecnologia (IIT), Milan, Italy.

Mattia Dalsass (M)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.

Marco Filipuzzi (M)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.

Alessia Loffreda (A)

Experimental Imaging Center, IRCCS San Raffaele Scientific Institute, Milan, Italy.

Alessandro Verrecchia (A)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.

Virginia Castillo Cano (V)

Peptomyc S.L., Barcelona, Spain.
Vall d'Hebron Institute of Oncology (VHIO), Edifici Cellex, Barcelona, Spain.

Hugo Thabussot (H)

Peptomyc S.L., Barcelona, Spain.

Mirko Doni (M)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.

Marco J Morelli (MJ)

Center for Genomic Science of IIT@SEMM, Fondazione Istituto Italiano di Tecnologia (IIT), Milan, Italy.

Laura Soucek (L)

Peptomyc S.L., Barcelona, Spain.
Vall d'Hebron Institute of Oncology (VHIO), Edifici Cellex, Barcelona, Spain.
Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain.
Department of Biochemistry and Molecular Biology, Universitat Autònoma de Barcelona, Bellaterra, Spain.

Theresia Kress (T)

Center for Genomic Science of IIT@SEMM, Fondazione Istituto Italiano di Tecnologia (IIT), Milan, Italy.

Davide Mazza (D)

Experimental Imaging Center, IRCCS San Raffaele Scientific Institute, Milan, Italy.

Marina Mapelli (M)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.

Marie-Eve Beaulieu (ME)

Peptomyc S.L., Barcelona, Spain.

Bruno Amati (B)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.

Arianna Sabò (A)

European Institute of Oncology (IEO) - IRCCS, Milan, Italy.

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Classifications MeSH