cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat
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 spe...
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| Format: | Article |
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Multidisciplinary Digital Publishing Institute
2023
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| Online Access: | https://hdl.handle.net/1721.1/150889 |
| _version_ | 1826196833177698304 |
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| author | Stalker, Leanne Backx, Alanna G. Tscherner, Allison K. Russell, Stewart J. Foster, Robert A. LaMarre, Jonathan |
| author2 | Massachusetts Institute of Technology. Division of Comparative Medicine |
| author_facet | Massachusetts Institute of Technology. Division of Comparative Medicine Stalker, Leanne Backx, Alanna G. Tscherner, Allison K. Russell, Stewart J. Foster, Robert A. LaMarre, Jonathan |
| author_sort | Stalker, Leanne |
| collection | MIT |
| description | 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 <i>PIWIL1</i> were cloned from feline testes cDNA. One isoform shows high homology to <i>PIWIL1</i> 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. |
| first_indexed | 2024-09-23T10:38:20Z |
| format | Article |
| id | mit-1721.1/150889 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T10:38:20Z |
| publishDate | 2023 |
| publisher | Multidisciplinary Digital Publishing Institute |
| record_format | dspace |
| spelling | mit-1721.1/1508892024-01-19T21:33:45Z cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat Stalker, Leanne Backx, Alanna G. Tscherner, Allison K. Russell, Stewart J. Foster, Robert A. LaMarre, Jonathan Massachusetts Institute of Technology. Division of Comparative Medicine Massachusetts Institute of Technology. Department of Biological Engineering 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 <i>PIWIL1</i> were cloned from feline testes cDNA. One isoform shows high homology to <i>PIWIL1</i> 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. 2023-06-09T15:06:10Z 2023-06-09T15:06:10Z 2023-05-26 2023-06-09T13:01:09Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/150889 International Journal of Molecular Sciences 24 (11): 9346 (2023) PUBLISHER_CC http://dx.doi.org/10.3390/ijms24119346 Creative Commons Attribution application/pdf Multidisciplinary Digital Publishing Institute Multidisciplinary Digital Publishing Institute |
| spellingShingle | Stalker, Leanne Backx, Alanna G. Tscherner, Allison K. Russell, Stewart J. Foster, Robert A. LaMarre, Jonathan cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat |
| title | cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat |
| title_full | cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat |
| title_fullStr | cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat |
| title_full_unstemmed | cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat |
| title_short | cDNA Cloning of Feline PIWIL1 and Evaluation of Expression in the Testis of the Domestic Cat |
| title_sort | cdna cloning of feline piwil1 and evaluation of expression in the testis of the domestic cat |
| url | https://hdl.handle.net/1721.1/150889 |
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