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Deleted in AZoospermia Associated Protein 1 (DAZAP1) is a ubiquitous hnRNP protein that has been implicated in RNA transcription, splicing, and translation. It is highly expressed in testes, predominantly in late stage spermatocytes and post-meiotic spermatids.  Dazap1  deficiency in mice results in growth retardation and spermatogenic arrest. The gene produces two major transcripts of 2.4 and 1.8 kb, designated  Dazap1 -L and  Dazap1 -S, respectively. Results of our previous RNA  in situ  hybridization and immunostaining suggested translational regulation of the  Dazap1  transcripts during spermatogenesis. The main objectives of the study were to determine the origin of the two  Dazap1  transcripts and to investigate whether they were similarly translated. Our Northern and 3′ RACE analyses showed that the two transcripts were generated through alternative polyadenylation. In mouse testes, the levels of both transcripts were low at postnatal day 12 (P12), increased significantly at P18, and reached maximum at P27. Sucrose gradient analyses showed that at P12 both transcripts were actively translated. Afterward, an increasing portion of  Dazap1 -S became associated with the translationally inactive mRNPs, and the translational repression was accompanied by an increase in the length of its poly(A) tail. A much smaller portion of  Dazap1 -L was also sequestered to mRNPs as testes matured, but there was no changes in its poly(A) tail length. Using RNA pull-down followed by mass spectrometry, we identified DAZL, a germ-cell specific translation regulator, as one of the proteins that bound to the 3′UTR region specific for  Dazap1 -L. We further showed that DAZL preferentially bound to  Dazap1- L in testis lysates and stimulated the translation of a reporter gene carrying  Dazap1 -L 3′UTR. In summary, our study shows that the translation of the two  Dazap1  transcripts is differentially regulated. It also provides a new example of translational repression associated with poly(A) tail elongation during spermatogenesis.

Differential Translation of Dazap1 Transcripts during Spermatogenesis