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bioRxiv · 10.64898/2026.06.23.733120

piRNAs from Y chromosomal protein coding, noncoding and endogenous retrovirus homologous repeat families regulate autosomal gene expression in mouse testis

Jesudasan, R.·Mukhoti, A.·Chaturvedi, A.·Tiwari, S.·Mishra, K.·Pranatharthi, A.·Praveena, N.·Alex, J.·Karunanithi, S.·Kumar, A.·Reddy, H.
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Abstract

BackgroundHeterochromatic long arm of mouse Y chromosome harbors the multicopy species-specific sequences Ssty, Sly, Asty and Orly that are transcribed in testis and have known functions in male fertility. Of these Ssty and Sly encode proteins - yet all the transcripts are not translated. To investigate the roles of these Y-heterochromatic transcripts further, we analyzed them. MethodsMice with 2/3rd deletion of the Y-chromosome (XYRIIIqdel) and its wild type (XYRIII) were used in this study. Bioinformatic approaches, small RNA northern blots, Electrophoretic Mobility Shift Assays, Luciferase reporter assays, dPCR analysis, RT-qPCR assays and western blotting techniques were used to identify piRNAs that regulate autosomal genes. ResultsWe demonstrate that the multicopy gene families from mouse Y-long arm generate piRNAs predominantly in testis. We observed sequences homologous to these piRNAs in the UTRs of a few autosomal genes, which are differentially expressed in the sperms of XYRIIIqdel mice. Furthermore, the Endogenous Retrovirus Element (ERV) LTR, found in the Orly1 transcript identified piRNAs in the database, showed homology to UTRs and associated genomic regions of a few autosomal genes. Orly1 showed a reduction in genomic copy number by digital PCR in XYRIIIqdel mice. One of the four autosomal genes containing the ERV segment in their UTRs, showed a differential testicular protein expression in the mutant mice. ConclusionsThus, we further elucidate that different classes of repeats from Y-chromosome regulate autosomal gene expression via piRNAs. Besides, this study also identified novel roles for a Y-derived ERV in autosomal gene regulation in testis.

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Jesudasan, R., Mukhoti, A., Chaturvedi, A., Tiwari, S., Mishra, K., Pranatharthi, A., Praveena, N., Alex, J., Karunanithi, S., Kumar, A., Reddy, H.. 2026-06-23. piRNAs from Y chromosomal protein coding, noncoding and endogenous retrovirus homologous repeat families regulate autosomal gene expression in mouse testis. https://doi.org/10.64898/2026.06.23.733120

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