cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat
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ijms-24-09346-v2.pdf
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Author(s)
Stalker, Leanne
Backx, Alanna G.
Tscherner, Allison K.
Russell, Stewart J.
Foster, Robert A.
LaMarre, Jonathan
Date Issued
May 26, 2023
Publisher
Multidisciplinary Digital Publishing Institute
Citation
International Journal of Molecular Sciences 24 (11): 9346 (2023)
Version
Final published version
Abstract
The PIWI clade of Argonaute proteins is essential for spermatogenesis in all species examined to date. This protein family binds specific classes of small non-coding RNAs known as PIWI-interacting RNAs (piRNAs) which together form piRNA-induced silencing complexes (piRISCs) that are recruited to specific RNA targets through sequence complementarity. These complexes facilitate gene silencing through endonuclease activity and guided recruitment of epigenetic silencing factors. PIWI proteins and piRNAs have been found to play multiple roles in the testis including the maintenance of genomic integrity through transposon silencing and facilitating the turnover of coding RNAs during spermatogenesis. In the present study, we report the first characterization of PIWIL1 in the male domestic cat, a mammalian system predicted to express four PIWI family members. Multiple transcript variants of PIWIL1 were cloned from feline testes cDNA. One isoform shows high homology to PIWIL1 from other mammals, however, the other has characteristics of a “slicer null” isoform, lacking the domain required for endonuclease activity. Expression of PIWIL1 in the male cat appears limited to the testis and correlates with sexual maturity. RNA-immunoprecipitation revealed that feline PIWIL1 binds small RNAs with an average size of 29 nt. Together, these data suggest that the domestic cat has two PIWIL1 isoforms expressed in the mature testis, at least one of which interacts with piRNAs.
MIT Department
Massachusetts Institute of Technology. Division of Comparative Medicine
Massachusetts Institute of Technology. Department of Biological Engineering
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DOI of Published Version
https://doi.org/10.3390/ijms24119346
