MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 Expression
Paternally-inherited loss-of-function mutations in makorin ring finger protein 3 gene (MKRN3) underlie central precocious puberty. To investigate the puberty-related mechanism(s) of MKRN3 in humans, we generated two distinct bi-allelic MKRN3 knock-out human pluripotent stem cell lines, Del 1 and Del...
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doaj-3d6a009648c84c70bc979e8d8fa6da7f2020-11-25T00:41:57ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922019-02-011010.3389/fendo.2019.00048430451MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 ExpressionVenkatram Yellapragada0Venkatram Yellapragada1Xiaonan Liu2Xiaonan Liu3Carina Lund4Carina Lund5Johanna Känsäkoski6Kristiina Pulli7Kristiina Pulli8Sanna Vuoristo9Karolina Lundin10Timo Tuuri11Markku Varjosalo12Markku Varjosalo13Taneli Raivio14Taneli Raivio15Taneli Raivio16Stem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Helsinki, FinlandDepartment of Physiology, Faculty of Medicine, University of Helsinki, Helsinki, FinlandMolecular Systems Biology Research Group, Institute of Biotechnology & HiLIFE, University of Helsinki, Helsinki, FinlandProteomics Unit, Institute of Biotechnology, University of Helsinki, Helsinki, FinlandStem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Helsinki, FinlandDepartment of Physiology, Faculty of Medicine, University of Helsinki, Helsinki, FinlandDepartment of Physiology, Faculty of Medicine, University of Helsinki, Helsinki, FinlandStem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Helsinki, FinlandDepartment of Physiology, Faculty of Medicine, University of Helsinki, Helsinki, FinlandDepartment of Obstetrics and Gynecology, Helsinki University Hospital, HUH, Helsinki, FinlandDepartment of Obstetrics and Gynecology, Helsinki University Hospital, HUH, Helsinki, FinlandDepartment of Obstetrics and Gynecology, Helsinki University Hospital, HUH, Helsinki, FinlandMolecular Systems Biology Research Group, Institute of Biotechnology & HiLIFE, University of Helsinki, Helsinki, FinlandProteomics Unit, Institute of Biotechnology, University of Helsinki, Helsinki, FinlandStem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Helsinki, FinlandDepartment of Physiology, Faculty of Medicine, University of Helsinki, Helsinki, FinlandNew Children's Hospital, Pediatric Research Center, Helsinki University Hospital, HUH, Helsinki, FinlandPaternally-inherited loss-of-function mutations in makorin ring finger protein 3 gene (MKRN3) underlie central precocious puberty. To investigate the puberty-related mechanism(s) of MKRN3 in humans, we generated two distinct bi-allelic MKRN3 knock-out human pluripotent stem cell lines, Del 1 and Del 2, and differentiated them into GNRH1-expressing neurons. Both Del 1 and Del 2 clones could be differentiated into neuronal progenitors and GNRH1-expressing neurons, however, the relative expression of GNRH1 did not differ from wild type cells (P = NS). Subsequently, we investigated stable and dynamic protein-protein interaction (PPI) partners of MKRN3 by stably expressing it in HEK cells followed by mass spectrometry analyses. We found 81 high-confidence novel protein interaction partners, which are implicated in cellular processes such as insulin signaling, RNA metabolism and cell-cell adhesion. Of the identified interactors, 20 have been previously implicated in puberty timing. In conclusion, our stem cell model for generation of GNRH1-expressing neurons did not offer mechanistic insight for the role of MKRN3 in puberty initiation. The PPI data, however, indicate that MKRN3 may regulate puberty by interacting with other puberty-related proteins. Further studies are required to elucidate the possible mechanisms and outcomes of these interactions.https://www.frontiersin.org/article/10.3389/fendo.2019.00048/fullMKRN3puberty timingprotein interactionGNRH1CRISPR/Cas9 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Venkatram Yellapragada Venkatram Yellapragada Xiaonan Liu Xiaonan Liu Carina Lund Carina Lund Johanna Känsäkoski Kristiina Pulli Kristiina Pulli Sanna Vuoristo Karolina Lundin Timo Tuuri Markku Varjosalo Markku Varjosalo Taneli Raivio Taneli Raivio Taneli Raivio |
spellingShingle |
Venkatram Yellapragada Venkatram Yellapragada Xiaonan Liu Xiaonan Liu Carina Lund Carina Lund Johanna Känsäkoski Kristiina Pulli Kristiina Pulli Sanna Vuoristo Karolina Lundin Timo Tuuri Markku Varjosalo Markku Varjosalo Taneli Raivio Taneli Raivio Taneli Raivio MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 Expression Frontiers in Endocrinology MKRN3 puberty timing protein interaction GNRH1 CRISPR/Cas9 |
author_facet |
Venkatram Yellapragada Venkatram Yellapragada Xiaonan Liu Xiaonan Liu Carina Lund Carina Lund Johanna Känsäkoski Kristiina Pulli Kristiina Pulli Sanna Vuoristo Karolina Lundin Timo Tuuri Markku Varjosalo Markku Varjosalo Taneli Raivio Taneli Raivio Taneli Raivio |
author_sort |
Venkatram Yellapragada |
title |
MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 Expression |
title_short |
MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 Expression |
title_full |
MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 Expression |
title_fullStr |
MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 Expression |
title_full_unstemmed |
MKRN3 Interacts With Several Proteins Implicated in Puberty Timing but Does Not Influence GNRH1 Expression |
title_sort |
mkrn3 interacts with several proteins implicated in puberty timing but does not influence gnrh1 expression |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Endocrinology |
issn |
1664-2392 |
publishDate |
2019-02-01 |
description |
Paternally-inherited loss-of-function mutations in makorin ring finger protein 3 gene (MKRN3) underlie central precocious puberty. To investigate the puberty-related mechanism(s) of MKRN3 in humans, we generated two distinct bi-allelic MKRN3 knock-out human pluripotent stem cell lines, Del 1 and Del 2, and differentiated them into GNRH1-expressing neurons. Both Del 1 and Del 2 clones could be differentiated into neuronal progenitors and GNRH1-expressing neurons, however, the relative expression of GNRH1 did not differ from wild type cells (P = NS). Subsequently, we investigated stable and dynamic protein-protein interaction (PPI) partners of MKRN3 by stably expressing it in HEK cells followed by mass spectrometry analyses. We found 81 high-confidence novel protein interaction partners, which are implicated in cellular processes such as insulin signaling, RNA metabolism and cell-cell adhesion. Of the identified interactors, 20 have been previously implicated in puberty timing. In conclusion, our stem cell model for generation of GNRH1-expressing neurons did not offer mechanistic insight for the role of MKRN3 in puberty initiation. The PPI data, however, indicate that MKRN3 may regulate puberty by interacting with other puberty-related proteins. Further studies are required to elucidate the possible mechanisms and outcomes of these interactions. |
topic |
MKRN3 puberty timing protein interaction GNRH1 CRISPR/Cas9 |
url |
https://www.frontiersin.org/article/10.3389/fendo.2019.00048/full |
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