scholarly journals CDK2 kinase activity is a regulator of male germ cell fate

2019 ◽  
Author(s):  
Priti Singh ◽  
Ravi K. Patel ◽  
Nathan Palmer ◽  
Jennifer K. Grenier ◽  
Darius Paduch ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Fate ◽  
Kinase Activity ◽  
Phosphorylation Site ◽  
Male Germ Cell ◽  
Cyclin Dependent Kinase ◽  
Rna Seq ◽  
Temporal Regulation ◽  
Juvenile Males

ABSTRACTThe ability of men to remain fertile throughout their lives depends upon establishment of a spermatogonial stem cell (SSC) pool from gonocyte progenitors, and also maintaining the proper balance between SSC renewal and spermatogenic differentiation throughout life. Depletion of SSCs causes infertility with a Sertoli Cell Only Syndrome (SCOS) phenotype. We previously created a mouse strain in which an inhibitory phosphorylation site (Tyr15) of Cyclin-dependent kinase 2 (Cdk2) was altered. Juvenile males homozygous for this allele (Cdk2Y15S) initiate the first round of spermatogenesis, which originates from prospermatogonia, but meiocytes arrest due to chromosomal defects resembling those in Cdk2-/- mice. Subsequent waves of spermatogonial differentiation and meiosis were largely absent, leading to an SCOS-like phenotype. Here, we demonstrate that Cdk2Y15S/Y15S mice possess mitotically active GFRa1+ SSC-like cells, but they are impaired in their ability to differentiate. Marker analysis and single cell RNA-seq revealed defective differentiation of gonocytes into SSCs. Biochemical and genetic data demonstrated that Cdk2Y15S is a gain-of-function allele causing deregulated kinase activity, and its phenotypic effects could be reversed by mutating the Thr160 positive regulatory site in cis. These results demonstrate that precise temporal regulation of CDK2 activity in male germ cell development and in the cell cycle is critical for long-term spermatogenic homeostasis.

scholarly journals CDK2 kinase activity is a regulator of male germ cell fate

Development ◽  
2019 ◽  
Vol 146 (21) ◽  
pp. dev180273 ◽  
Author(s):  
Priti Singh ◽  
Ravi K. Patel ◽  
Nathan Palmer ◽  
Jennifer K. Grenier ◽  
Darius Paduch ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Fate ◽  
Kinase Activity ◽  
Male Germ Cell

Long-term results of a combination of paclitaxel, cisplatin and gemcitabine for salvage therapy in male germ-cell tumours

2009 ◽  
Vol 104 (3) ◽  
pp. 340-346 ◽  
Author(s):  
Nicola Nicolai ◽  
Andrea Necchi ◽  
Luca Gianni ◽  
Luigi Piva ◽  
Davide Biasoni ◽  
...  
Keyword(s):  
Germ Cell ◽  
Salvage Therapy ◽  
Male Germ Cell ◽  
Long Term Results ◽  
Germ Cell Tumours

scholarly journals Modified cisplatin, etoposide (or vinblastine) and ifosfamide salvage therapy for male germ-cell tumors. Long-term results

1992 ◽  
Vol 3 (3) ◽  
pp. 211-216 ◽  
Author(s):  
G. Pizzocaro ◽  
R. Salvioni ◽  
L. Piva ◽  
M. Faustini ◽  
N. Nicolai ◽  
...  
Keyword(s):  
Germ Cell ◽  
Salvage Therapy ◽  
Germ Cell Tumors ◽  
Male Germ Cell ◽  
Long Term Results

Porcine nuclear transfer using somatic donor cells altered to express male germ cell function

2009 ◽  
Vol 21 (7) ◽  
pp. 882 ◽  
Author(s):  
Sangho Roh ◽  
Hye-Yeon Choi ◽  
Sang Kyu Park ◽  
Cheolhee Won ◽  
Bong-Woo Kim ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cell Fate ◽  
Nuclear Transfer ◽  
Cell Function ◽  
Control Cell ◽  
Donor Cell ◽  
Male Germ Cell ◽  
Cell Group ◽  
Cell Extracts

Recent studies reported that the direct transformation of one differentiated somatic cell type into another is possible. In the present study, we were able to modulate the cell fate of somatic cells to take on male germ cell function by introducing cell extracts derived from porcine testis tissue. Fibroblasts were treated with streptolysin O, which reversibly permeabilises the plasma membrane, and incubated with testis extracts. Our results showed that the testis extracts (TE) could activate expression of male germ cell-specific genes, implying that TE can provide regulatory components required for altering the cell fate of fibroblasts. Male germ cell function was sustained for more than 10 days after the introduction of TE. In addition, a single TE-treated cell was injected directly into the cytoplasm of in vitro-matured porcine oocytes. The rate of blastocyst formation was significantly higher in the TE-treated nuclear donor cell group than in the control cell group. The expression level of Nanog, Sox9 and Eomes was drastically increased when altered cells were used as donor nuclei. Our results suggest that TE can be used to alter the cell fate of fibroblasts to express male germ cell function and improve the developmental efficiency of the nuclear transfer porcine embryos.

scholarly journals Proliferation and cell fate establishment during Arabidopsis male gametogenesis depends on the Retinoblastoma protein

2009 ◽  
Vol 106 (17) ◽  
pp. 7257-7262 ◽  
Author(s):  
Zhong Chen ◽  
Said Hafidh ◽  
Shi Hui Poh ◽  
David Twell ◽  
Frederic Berger
Keyword(s):  
Cell Proliferation ◽  
Germ Cell ◽  
Cell Fate ◽  
Vegetative Cell ◽  
Germ Line ◽  
Sperm Cells ◽  
Cyclin Dependent Kinase ◽  
Rb Protein ◽  
Male Gametogenesis ◽  
Male Germline

The Retinoblastoma (Rb) protein is a conserved repressor of cell proliferation. In animals and plants, deregulation of Rb protein causes hyperproliferation and perturbs cell differentiation to various degrees. However, the primary developmental impact of the loss of Rb protein has remained unclear. In this study we investigated the direct consequences of Rb protein knockout in the Arabidopsis male germline using cytological and molecular markers. The Arabidopsis germ line derives from the unequal division of the microspore, producing a small germ cell and a large terminally differentiated vegetative cell. A single division of the germ cell produces the 2 sperm cells. We observed that the loss of Rb protein does not have a major impact on microspore division but causes limited hyperproliferation of the vegetative cell and, to a lesser degree, of the sperm cells. In addition, cell fate is perturbed in a fraction of Rb-defective vegetative cells. These defects are rescued by preventing cell proliferation arising from down-regulation of cyclin-dependent kinase A1. Our results indicate that hyperproliferation caused by the loss of Rb protein prevents or delays cell determination during plant male gametogenesis, providing further evidence for a direct link between fate determination and cell proliferation.

scholarly journals BRCA1 Is Phosphorylated at Serine 1497 In Vivo at a Cyclin-Dependent Kinase 2 Phosphorylation Site

1999 ◽  
Vol 19 (7) ◽  
pp. 4843-4854 ◽  
Author(s):  
Heinz Ruffner ◽  
Wei Jiang ◽  
A. Grey Craig ◽  
Tony Hunter ◽  
Inder M. Verma
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Progression ◽  
Kinase Activity ◽  
Phosphorylation Site ◽  
Cyclin A ◽  
Dominant Negative ◽  
Protein Kinase Activity ◽  
Cyclin Dependent Kinase

ABSTRACT BRCA1 is a cell cycle-regulated nuclear protein that is phosphorylated mainly on serine and to a lesser extent on threonine residues. Changes in phosphorylation occur in response to cell cycle progression and DNA damage. Specifically, BRCA1 undergoes hyperphosphorylation during late G1 and S phases of the cell cycle. Here we report that BRCA1 is phosphorylated in vivo at serine 1497 (S1497), which is part of a cyclin-dependent kinase (CDK) consensus site. S1497 can be phosphorylated in vitro by CDK2-cyclin A or E. BRCA1 coimmunoprecipitates with an endogenous serine-threonine protein kinase activity that phosphorylates S1497 in vitro. This cellular kinase activity is sensitive to transfection of a dominant negative form of CDK2 as well as the application of the CDK inhibitors p21 and butyrolactone I but not p16. Furthermore, BRCA1 coimmunoprecipitates with CDK2 and cyclin A. These results suggest that the endogenous kinase activity is composed of CDK2-cyclin complexes, at least in part, concordant with the G1/S-specific increase in BRCA1 phosphorylation.

747 MODIFIED CISPLATIN, ETOPOSIDE AND IFOSFAMIDE (PEI) SALVAGE THERAPY FOR MALE GERM-CELL TUMORS (GCT). LONG-TERM EFFICACY AND SAFETY OUTCOMES

2013 ◽  
Vol 189 (4S) ◽  
Author(s):  
Andrea Necchi ◽  
Luigi Mariani ◽  
Patrizia Giannatempo ◽  
Nicola Nicolai ◽  
Daniele Raggi ◽  
...  
Keyword(s):  
Germ Cell ◽  
Salvage Therapy ◽  
Germ Cell Tumors ◽  
Male Germ Cell ◽  
Efficacy And Safety ◽  
Term Efficacy ◽  
Safety Outcomes

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

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 984
Author(s):  
Maciej J. Smialek ◽  
Erkut Ilaslan ◽  
Marcin P. Sajek ◽  
Aleksandra Swiercz ◽  
Damian M. Janecki ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Germ Cell ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Cell Model ◽  
Human Reproduction ◽  
Male Germ Cell ◽  
Binding Motif ◽  
Rna Seq ◽  
Human Male

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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