Context-dependent functional compensation between Ythdf m6A reader proteins.
The N6-methyladenosine (m6A) modification is the most prevalent post-transcriptional mRNA modification, regulating mRNA decay and splicing. It plays a major role during normal development, differentiation, and disease progression. The modification is regulated by a set of writer, eraser, and reader proteins. The YTH domain family of proteins consists of three homologous m6A-binding proteins, Ythdf1, Ythdf2, and Ythdf3, which were suggested to have different cellular functions. However, their sequence similarity and their tendency to bind the same targets suggest that they may have overlapping roles. We systematically knocked out (KO) the Mettl3 writer, each of the Ythdf readers, and the three readers together (triple-KO). We then estimated the effect in vivo in mouse gametogenesis, postnatal viability, and in vitro in mouse embryonic stem cells (mESCs). In gametogenesis, Mettl3-KO severity is increased as the deletion occurs earlier in the process, and Ythdf2 has a dominant role that cannot be compensated by Ythdf1 or Ythdf3, due to differences in readers' expression pattern across different cell types, both in quantity and in spatial location. Knocking out the three readers together and systematically testing viable offspring genotypes revealed a redundancy in the readers' role during early development that is Ythdf1/2/3 gene dosage-dependent. Finally, in mESCs there is compensation between the three Ythdf reader proteins, since the resistance to differentiate and the significant effect on mRNA decay occur only in the triple-KO cells and not in the single KOs. Thus, we suggest a new model for the Ythdf readers function, in which there is profound dosage-dependent redundancy when all three readers are equivalently coexpressed in the same cell types.
['Lasman|Lior|L|', 'Krupalnik|Vladislav|V|', 'Viukov|Sergey|S|', 'Mor|Nofar|N|', 'Aguilera-Castrejon|Alejandro|A|', 'Schneir|Dan|D|', 'Bayerl|Jonathan|J|', 'Mizrahi|Orel|O|', 'Peles|Shani|S|', 'Tawil|Shadi|S|', 'Sathe|Shashank|S|', 'Nachshon|Aharon|A|', 'Shani|Tom|T|', 'Zerbib|Mirie|M|', 'Kilimnik|Itay|I|', 'Aigner|Stefan|S|', 'Shankar|Archana|A|', 'Mueller|Jasmine R|JR|', 'Schwartz|Schraga|S|', 'Stern-Ginossar|Noam|N|0000-0003-2042-9974', 'Yeo|Gene W|GW|', 'Geula|Shay|S|', 'Novershtern|Noa|N|', 'Hanna|Jacob H|JH|0000-0003-2042-9974']
D000818:Animals D002460:Cell Line D004303:Dosage Compensation, Genetic D053595:Embryonic Stem Cells D005298:Fertility D005718:Gametogenesis D017353:Gene Deletion D020869:Gene Expression Profiling D018507:Gene Expression Regulation, Developmental D008780:Methyltransferases D051379:Mice D018345:Mice, Knockout D016601:RNA-Binding Proteins
m6A stem cells RNA methylation