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Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model

Mammalian Pumilio (PUM) proteins are sequence-specific, RNA-binding proteins (RBPs) with wide-ranging roles. They are involved in germ cell development, which has functional implications in development and fertility. Although human PUM1 and PUM2 are closely related to each other and recognize the sa...

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Autores principales: Smialek, Maciej J., Ilaslan, Erkut, Sajek, Marcin P., Swiercz, Aleksandra, Janecki, Damian M., Kusz-Zamelczyk, Kamila, Wozniak, Tomasz, Kotecki, Maciej, Handschuh, Luiza, Figlerowicz, Marek, Jaruzelska, Jadwiga
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7226987/
https://www.ncbi.nlm.nih.gov/pubmed/32316190
http://dx.doi.org/10.3390/cells9040984
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author Smialek, Maciej J.
Ilaslan, Erkut
Sajek, Marcin P.
Swiercz, Aleksandra
Janecki, Damian M.
Kusz-Zamelczyk, Kamila
Wozniak, Tomasz
Kotecki, Maciej
Handschuh, Luiza
Figlerowicz, Marek
Jaruzelska, Jadwiga
author_facet Smialek, Maciej J.
Ilaslan, Erkut
Sajek, Marcin P.
Swiercz, Aleksandra
Janecki, Damian M.
Kusz-Zamelczyk, Kamila
Wozniak, Tomasz
Kotecki, Maciej
Handschuh, Luiza
Figlerowicz, Marek
Jaruzelska, Jadwiga
author_sort Smialek, Maciej J.
collection PubMed
description Mammalian Pumilio (PUM) proteins are sequence-specific, RNA-binding proteins (RBPs) with wide-ranging roles. They are involved in germ cell development, which has functional implications in development and fertility. Although human PUM1 and PUM2 are closely related to each other and recognize the same RNA binding motif, there is some evidence for functional diversity. To address that problem, first we used RIP-Seq and RNA-Seq approaches, and identified mRNA pools regulated by PUM1 and PUM2 proteins in the TCam-2 cell line, a human male germ cell model. Second, applying global mass spectrometry-based profiling, we identified distinct PUM1- and PUM2-interacting putative protein cofactors, most of them involved in RNA processing. Third, combinatorial analysis of RIP and RNA-Seq, mass spectrometry, and RNA motif enrichment analysis revealed that PUM1 and PUM2 form partially varied RNP-regulatory networks (RNA regulons), which indicate different roles in human reproduction and testicular tumorigenesis. Altogether, this work proposes that protein paralogues with very similar and evolutionary highly conserved functional domains may play divergent roles in the cell by combining with different sets of protein cofactors. Our findings highlight the versatility of PUM paralogue-based post-transcriptional regulation, offering insight into the mechanisms underlying their diverse biological roles and diseases resulting from their dysfunction.
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spelling pubmed-72269872020-05-18 Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model Smialek, Maciej J. Ilaslan, Erkut Sajek, Marcin P. Swiercz, Aleksandra Janecki, Damian M. Kusz-Zamelczyk, Kamila Wozniak, Tomasz Kotecki, Maciej Handschuh, Luiza Figlerowicz, Marek Jaruzelska, Jadwiga Cells Article Mammalian Pumilio (PUM) proteins are sequence-specific, RNA-binding proteins (RBPs) with wide-ranging roles. They are involved in germ cell development, which has functional implications in development and fertility. Although human PUM1 and PUM2 are closely related to each other and recognize the same RNA binding motif, there is some evidence for functional diversity. To address that problem, first we used RIP-Seq and RNA-Seq approaches, and identified mRNA pools regulated by PUM1 and PUM2 proteins in the TCam-2 cell line, a human male germ cell model. Second, applying global mass spectrometry-based profiling, we identified distinct PUM1- and PUM2-interacting putative protein cofactors, most of them involved in RNA processing. Third, combinatorial analysis of RIP and RNA-Seq, mass spectrometry, and RNA motif enrichment analysis revealed that PUM1 and PUM2 form partially varied RNP-regulatory networks (RNA regulons), which indicate different roles in human reproduction and testicular tumorigenesis. Altogether, this work proposes that protein paralogues with very similar and evolutionary highly conserved functional domains may play divergent roles in the cell by combining with different sets of protein cofactors. Our findings highlight the versatility of PUM paralogue-based post-transcriptional regulation, offering insight into the mechanisms underlying their diverse biological roles and diseases resulting from their dysfunction. MDPI 2020-04-16 /pmc/articles/PMC7226987/ /pubmed/32316190 http://dx.doi.org/10.3390/cells9040984 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Smialek, Maciej J.
Ilaslan, Erkut
Sajek, Marcin P.
Swiercz, Aleksandra
Janecki, Damian M.
Kusz-Zamelczyk, Kamila
Wozniak, Tomasz
Kotecki, Maciej
Handschuh, Luiza
Figlerowicz, Marek
Jaruzelska, Jadwiga
Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model
title Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model
title_full Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model
title_fullStr Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model
title_full_unstemmed Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model
title_short Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model
title_sort characterization of rnp networks of pum1 and pum2 post-transcriptional regulators in tcam-2 cells, a human male germ cell model
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7226987/
https://www.ncbi.nlm.nih.gov/pubmed/32316190
http://dx.doi.org/10.3390/cells9040984
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