Contact inhibition of proliferation is accompanied by expression of the PHF10D subunit of the chromatin remodeling complex PBAF in mouse and human cell lines

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Abstract

PHF10 is a subunit of the PBAF complex, which regulates the expression of many genes in developing and maturing organisms. PHF10 has four isoforms that differ in domain structure. The PHF10A isoform, containing a DPF domain at the C-terminus and 46 amino acids at the N-terminus, is necessary for the expression of gene proliferation; the functions of the other isoforms are less studied. In this work, we have established that upon contact inhibition of mouse and human cell proliferation caused by the establishment of a tight junction and adherence junction between cells, the expression of the PHF10A isoform stops and instead the PHF10D isoform is expressed, which does not contain DPF-domain and N-terminal sequence. The function of the PHF10D isoform may be associated with the establishment of intercellular contacts.

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About the authors

Yu. P. Simonov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: so2615nat@gmail.com

Department of Transcription Factors

Russian Federation, Moscow

V. V. Tatarskiy

Institute of Gene Biology, Russian Academy of Sciences

Email: so2615nat@gmail.com
Russian Federation, Moscow

S. G. Georgieva

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: so2615nat@gmail.com

academician of the RAS, Department of Transcription Factors

Russian Federation, Moscow

N. V. Soshnikova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences; Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Author for correspondence.
Email: so2615nat@gmail.com

Department of Transcription Factors

Russian Federation, Moscow; Moscow

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2. Fig. 1. (a) Schematic representation of the PBAF complex. Different subunits are indicated in different colors and the modules of the complex are indicated (adapted from [1]). (b) The domain organization of the PHF10 isoforms. Various domains are indicated. Green, blue and orange indicate the sequences that distinguish the isoforms.

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3. Fig. 2. (a) Mouse embryonic fibroblasts of MEF 1, 3, 4 and 5 days after passage on a 60 mm Petri dish. (b) Western blotting isoforms of Phf10 (indicated on the right in the upper panel), subunits of the PBAF complex: Baf180, Brg1, Baf155 in MEF lysates selected in different densities. Staining with antibodies to b- tubulin was used as a control of application. A change in the expression of Cyclin D1 (CyclinD1) and P27 characterizes the ability of cells to proliferate. (c) Changes in the mRNA level of Phf10 isoforms containing the DPF domain (Phf10P = Phf10A + Phf10B) and containing a PDSM motif (Phf10-S = Phf10C + Phf10D) at the C-terminus. The Y-axis shows the level change RNA is relative to control when normalized to the expression level of the RPLP0 gene. In all experiments, the measurements were carried out in 3 repetitions and presented as an average ± standard deviation . * – p < 0.005 comparison with control (single-factor analysis of variance with the criterion of multiple comparisons of Dunnet).

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4. Fig. 3. Western blotting of PHF10 isoforms (shown on the right in the upper panel) in lysates of RKO cells selected in different densities. Staining with antibodies to b-tubulin was used as a control of application. Changes in the expression of MYC and P27 characterize the ability of cells to proliferate.

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