Handling Editor: Haruhiko Koseki
ABSTRACT
NANOS2 and NANOS3 are evolutionarily conserved RNA-binding proteins involved in murine germ cell development. NANOS3 is required for protection from apoptosis during migration and gonadal colonization in both sexes, whereas NANOS2 is male-specific and required for the male-type differentiation of germ cells. Ectopic NANOS2 rescues the functions of NANOS3, but NANOS3 cannot rescue NANOS2 function, even though its expression is upregulated in Nanos2-null conditions. It is unknown why NANOS3 cannot rescue NANOS2 function and it is unclear whether NANOS3 plays any role in male germ cell differentiation. To address these questions, we made conditional Nanos3/Nanos2 knockout mice and chimeric mice expressing chimeric NANOS proteins. Conditional double knockout of Nanos2 and Nanos3 led to the rapid loss of germ cells, and in vivo and in vitro experiments revealed that DND1 and NANOS2 binding is dependent on the specific NANOS2 zinc-finger structure. Moreover, murine NANOS3 failed to bind CNOT1, an interactor of NANOS2 at its N-terminal. Collectively, our study suggests that the inability of NANOS3 to rescue NANOS2 function is due to poor DND1 recruitment and CNOT1 binding.
Footnotes
Competing interests
The authors declare no competing or financial interests.
Author contributions
Conceptualization: D.W., Y.S.; Methodology: D.W., M.K., Y.S.; Validation: D.W.; Formal analysis: D.W.; Investigation: D.W.; Resources: M.K., Y.S.; Writing - original draft: D.W., Y.S.; Writing - review & editing: D.W., Y.S.; Visualization: D.W., Y.S.; Supervision: Y.S.; Project administration: Y.S.; Funding acquisition: Y.S.
Funding
This work was supported by the Japan Society for the Promotion of Science KAKENHI grant numbers 16H06279 and 17H06166.
Data availability
Single-cell RNA-seq data have been deposited into the DNA Data Bank of Japan (DDBJ) under accession number DRA011172.
Supplementary information
Supplementary information available online at https://dev.biologists.org/lookup/doi/10.1242/dev.191916.supplemental
- Received April 22, 2020.
- Accepted November 4, 2020.
Log in using your username and password
Log in through your institution
Pay Per Article - You may access this article (from the computer you are currently using) for 1 day for US$30.00 .
Regain Access - You can regain access to a recent Pay per Article purchase if your access period has not yet expired.