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| Status |
Public on Sep 11, 2012 |
| Title |
Alkbh1/Tzfp Repress a Non-Repeat piRNA Cluster in Pachytene Spermatocytes |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
Non-coding RNA profiling by high throughput sequencing
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| Summary |
Piwi proteins and Piwi-interacting small RNAs (piRNAs) have known functions in transposon silencing in the male germline of fetal and newborn mice. Both are also necessary for spermatogenesis in adult testes, however, their function here remains a mystery. Here, we use germ cell isolations and small RNA sequencing to show that most piRNAs in meiotic spermatocytes originate from clusters in intergenic non-repeat regions of DNA. The regulation of these piRNA clusters, including the processing of the precursor transcripts into individual piRNAs, is accomplished through mostly unknown processes. We present evidence for a regulatory mechanism for one such cluster, named cluster 1082B, located on chromosome 7 in the mouse genome, containing 788 unique piRNAs. The precursor transcript and individual piRNAs within the cluster are repressed by the Alkbh1 dioxygenase and the transcription repressor Tzfp, which are believed to be interaction partners in testis. We observe more than a thousand-fold upregulation of individual piRNAs in pachytene spermatocytes isolated from Alkbh1-/- and TzfpGTi/GTi testes. Repression is further supported by the identification of a 10 bp Tzfp recognition sequence contained within the precursor transcript. Downregulation of long interspersed elements 1 (LINE1) and intracisternal A-particle (IAP) transcripts in the Alkbh1-/- and TzfpGTi/GTi testes leads us to propose a potential role for the 1082B-encoded piRNAs in transposon silencing.
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| Overall design |
Characterization of small RNAs in mouse pachytene spermatocytes for wild-type (WT) and Alkbh1-/- and TzfpGTi/GTi, and mRNA in mouse pachytene spermatocytes for wild-type (WT) and Alkbh1-/-
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| Contributor(s) |
Nordstrand LM, Furu K, Paulsen J, Rognes T, Klungland A |
| Citation(s) |
22965116 |
| |
| Submission date |
Apr 10, 2012 |
| Last update date |
May 15, 2019 |
| Contact name |
Jonas Paulsen |
| Organization name |
Oslo University Hospital
|
| Department |
Radium Hospital
|
| Lab |
Institute for Medical Informatics
|
| Street address |
Montebello
|
| City |
Oslo |
| State/province |
Foreign |
| ZIP/Postal code |
N-0310 |
| Country |
Norway |
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| Platforms (1) |
| GPL11002 |
Illumina Genome Analyzer IIx (Mus musculus) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA158427 |
| SRA |
SRP012147 |
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