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Open AccessArticle

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

1
Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479 Poznan, Poland
2
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland
3
Institute of Computing Science, Poznan University of Technology, Piotrowo 2, 60-965 Poznan, Poland
4
Department of Developmental, Molecular and Chemical Biology, Tufts University Medical School, 136 Harrison Ave, Boston, MA 02111, USA
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Cells 2020, 9(4), 984; https://doi.org/10.3390/cells9040984
Received: 22 March 2020 / Revised: 9 April 2020 / Accepted: 13 April 2020 / Published: 16 April 2020
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. View Full-Text
Keywords: 3′UTR; human infertility; mass spectrometry; post-transcriptional gene regulation; ribonucleoprotein complexes; RNA-binding proteins (RBPs); testicular germ cell tumors 3′UTR; human infertility; mass spectrometry; post-transcriptional gene regulation; ribonucleoprotein complexes; RNA-binding proteins (RBPs); testicular germ cell tumors
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Smialek, M.J.; Ilaslan, E.; Sajek, M.P.; Swiercz, A.; Janecki, D.M.; Kusz-Zamelczyk, K.; Wozniak, T.; Kotecki, M.; Handschuh, L.; Figlerowicz, M.; Jaruzelska, J. Characterization of RNP Networks of PUM1 and PUM2 Post-Transcriptional Regulators in TCam-2 Cells, a Human Male Germ Cell Model. Cells 2020, 9, 984.

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