N6-Methyladenosine detected in RNA of testicular germ cell tumors is controlled by METTL3, ALKBH5, YTHDC1/F1/F2, and HNRNPC as writers, erasers, and readers.
Adenosine
/ analogs & derivatives
AlkB Homolog 5, RNA Demethylase
/ metabolism
Animals
Cell Line, Tumor
Heterogeneous-Nuclear Ribonucleoprotein Group C
/ metabolism
Humans
Male
Methyltransferases
/ metabolism
Mice
Neoplasms, Germ Cell and Embryonal
/ metabolism
Nerve Tissue Proteins
/ metabolism
RNA
/ metabolism
RNA Splicing Factors
/ metabolism
RNA-Binding Proteins
/ metabolism
Testicular Neoplasms
/ metabolism
DNA/RNA modification
N6-methyladenosine
eraser
germ cell tumor
reader
writer
Journal
Andrology
ISSN: 2047-2927
Titre abrégé: Andrology
Pays: England
ID NLM: 101585129
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
received:
21
12
2018
revised:
18
02
2019
accepted:
24
02
2019
pubmed:
25
3
2019
medline:
21
7
2020
entrez:
24
3
2019
Statut:
ppublish
Résumé
Type II testicular germ cell tumors (GCTs) arise from a common precursor lesion (germ cell neoplasia in situ) and are stratified into seminomas and non-seminomas, which differ considerably in morphology, gene expression, and epigenetic landscape. The N6-methyladenosine (6mA) epigenetic modification is the most abundant modification in mRNA and is also detectable in eukaryotic DNA. The functional role of 6mA is not fully understood, but 6mA residues may influence transcription by affecting splicing, miRNA processing, and mRNA stability. Additionally, the methyl group of 6mA destabilizes Watson-Crick base-pairing affecting RNA structure and protein binding. Here, we analyzed the presence of the 6mA epigenetic modification in germ cells and GCT tissues and cell lines. We screened for the presence of 6mA in DNA and RNA by immunohistochemistry, mass spectrometry or ELISA-based quantification assays. Additionally, expression of 6mA writer-, eraser- and reader-factors was analyzed by microarrays, qRT-PCR, western blotting and screening of public databases. We demonstrate that 6mA is detectable in RNA, but not DNA, of GCT cell lines and tissues, fibroblasts, and Sertoli cells as well as germ cells of different developmental stages. Based on expression analyses, our results suggest METTL3, ALKBH5, YTHDC1, YTHDF1, YTHDF2 and HNRNPC as main writers, erasers, and readers of the 6mA modification in GCTs. Owing to the lack of 6mA in DNA of GCTs, a functional role in regulating DNA transcription can be excluded. Interestingly, expression levels of 6mA regulators are comparable between tumor and normal tissues/cells, suggesting a similar mechanism of 6mA regulation in RNA. Finally, we demonstrate that 6mA levels in RNA increase upon differentiation of GCT cell lines, suggesting a role of 6mA in cell fate decisions. In summary, our data provide the starting point for further experiments deciphering the role of 6mA in the RNA of GCTs.
Sections du résumé
BACKGROUND
Type II testicular germ cell tumors (GCTs) arise from a common precursor lesion (germ cell neoplasia in situ) and are stratified into seminomas and non-seminomas, which differ considerably in morphology, gene expression, and epigenetic landscape. The N6-methyladenosine (6mA) epigenetic modification is the most abundant modification in mRNA and is also detectable in eukaryotic DNA. The functional role of 6mA is not fully understood, but 6mA residues may influence transcription by affecting splicing, miRNA processing, and mRNA stability. Additionally, the methyl group of 6mA destabilizes Watson-Crick base-pairing affecting RNA structure and protein binding.
OBJECTIVES
Here, we analyzed the presence of the 6mA epigenetic modification in germ cells and GCT tissues and cell lines.
MATERIALS AND METHODS
We screened for the presence of 6mA in DNA and RNA by immunohistochemistry, mass spectrometry or ELISA-based quantification assays. Additionally, expression of 6mA writer-, eraser- and reader-factors was analyzed by microarrays, qRT-PCR, western blotting and screening of public databases.
RESULTS
We demonstrate that 6mA is detectable in RNA, but not DNA, of GCT cell lines and tissues, fibroblasts, and Sertoli cells as well as germ cells of different developmental stages. Based on expression analyses, our results suggest METTL3, ALKBH5, YTHDC1, YTHDF1, YTHDF2 and HNRNPC as main writers, erasers, and readers of the 6mA modification in GCTs.
DISCUSSION
Owing to the lack of 6mA in DNA of GCTs, a functional role in regulating DNA transcription can be excluded. Interestingly, expression levels of 6mA regulators are comparable between tumor and normal tissues/cells, suggesting a similar mechanism of 6mA regulation in RNA. Finally, we demonstrate that 6mA levels in RNA increase upon differentiation of GCT cell lines, suggesting a role of 6mA in cell fate decisions.
CONCLUSION
In summary, our data provide the starting point for further experiments deciphering the role of 6mA in the RNA of GCTs.
Substances chimiques
HNRNPC protein, human
0
Heterogeneous-Nuclear Ribonucleoprotein Group C
0
Nerve Tissue Proteins
0
RNA Splicing Factors
0
RNA-Binding Proteins
0
YTHDC1 protein, human
0
YTHDF1 protein, human
0
YTHDF2 protein, human
0
RNA
63231-63-0
N-methyladenosine
CLE6G00625
ALKBH5 protein, human
EC 1.14.11.-
AlkB Homolog 5, RNA Demethylase
EC 1.14.11.-
Methyltransferases
EC 2.1.1.-
METTL3 protein, human
EC 2.1.1.62
Adenosine
K72T3FS567
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
498-506Subventions
Organisme : Deutsche Forschungsgemeinschaft
Pays : International
Informations de copyright
© 2019 American Society of Andrology and European Academy of Andrology.