Neuronal and muscle differentiation of mammalian cells is accompanied by a change of PHF10 isoform expression

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Abstract

The PBAF chromatin remodeling complex of the SWI/SNF family plays a critical role in the regulation of gene expression during tissue differentiation and organism development. The subunits of the PBAF complex have domains responsible for binding to N-terminal histone sequences. It determines the specificity of binding of the complex to chromatin. PHF10, a specific subunit of the PBAF complex, contains a DPF domain, which is a unique chromatin interaction domain. A PHF10 isoform that lacks the DPF domain is also present in vertebrate cells. This work shows that during neuronal and muscle differentiation of human and mouse cells, the expression of PHF10 isoforms changes: the form that does not have DPF replaces the form in which it is present. Replacement of PHF10 isoforms in the PBAF complex may affect its selectivity in the regulation of genes in differentiating cells.

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About the authors

D. O. Bayramova

Department of Transcription Factors, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: so2615@gmail.com
Russian Federation, Moscow; Moscow

A. M. Azieva

National Research Center "Kurchatov Institute"

Email: so2615@gmail.com
Russian Federation, Moscow

A. V. Feoktistov

Department of Transcription Factors, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: so2615@gmail.com
Russian Federation, Moscow; Moscow

S. G. Georgieva

Department of Transcription Factors, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: so2615@gmail.com

Academician of the RAS

Russian Federation, Moscow

N. V. Soshnikova

Department of Transcription Factors, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences; Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Author for correspondence.
Email: so2615@gmail.com
Russian Federation, Moscow; Moscow

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2. Fig. 1. (a) The scheme of PHF10 isoforms. The characteristic domains and motifs of the PHF10 isoforms are indicated. (b) The predicted structure of PHF10-Pl (Q8WUB8) according to the Alpha Fold database (https://alphafold.ebi.ac.uk/entry/Q8WUB8 ). (c) Expression of PHF10-P, PHF10-S isoforms and their ratio in various human tissues according to the GTEx database (The Genotype-Tissue Expression (GTEx) Project data). Along the ordinate axis of the TPM (the number of transcripts per million reads in the sample).

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3. Fig. 2. (a) and (b) Neuronal differentiation of human cells of the SH-SY5Y line (A) and mouse cells of the Neuro 2a line (b): left panel – visualization using negative contrast of the cell phenotype during ten days of differentiation; right panel – Western blotting of PHF10 isoforms without differentiation, in in the middle and at the end of differentiation. Isoforms are indicated on the right. The marker of molecular weights 55 and 70 kDa is shown on the left. Tubulin antibody staining was used as a control of application in both cases.

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4. Fig. 3. Western blotting of Phf10 isoforms in the process of myogenic differentiation of mouse cells of the line C2C12. Isoforms are indicated on the right. The marker of molecular weights - 55 and 70 kDa – is shown on the left. The coating with antibodies to the Gapgh protein was used as a control of application.

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