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RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Mus musculus

PROVIDER: GSE228454 | GEO | 2023/12/18

REPOSITORIES: GEO

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RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis.

Project description:Male germ cell development requires precise regulation of gene activity in a cell-type and stage-specific manner, with perturbations in gene expression during spermatogenesis associated with infertility. Here, we use steady-state, nascent and single-cell RNA sequencing strategies to comprehensively characterize gene expression across male germ cell populations, to dissect the mechanisms of gene control and provide new insights towards therapy. We discover a requirement for pausing of RNA Polymerase II (Pol II) at the earliest stages of sperm differentiation to establish the landscape of gene activity across development. Accordingly, genetic knockout of the Pol II pause-inducing factor NELF in immature germ cells blocks differentiation to spermatids. Further, we uncover unanticipated roles for Pol II pausing in the regulation of meiosis during spermatogenesis, with the presence of paused Pol II associated with double-strand break (DSB) formation, and disruption of meiotic gene expression and DSB repair in germ cells lacking NELF.

| S-EPMC10824759 | biostudies-literature

RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis.

Project description:Male germ cell development requires precise regulation of gene activity in a cell-type and stage-specific manner, with perturbations in gene expression during spermatogenesis associated with infertility. Here, we use steady-state, nascent and single-cell RNA sequencing strategies to comprehensively characterize gene expression across male germ cell populations, to dissect the mechanisms of gene control and provide new insights towards therapy. We discover a requirement for pausing of RNA Polymerase II (Pol II) at the earliest stages of sperm differentiation to establish the landscape of gene activity across development. Accordingly, genetic knockout of the Pol II pause-inducing factor NELF in immature germ cells blocks differentiation to mature spermatids. Further, we uncover unanticipated roles for Pol II pausing in the regulation of meiosis during spermatogenesis, with the presence of paused Pol II associated with double strand break formation by SPO11, and disruption of SPO11 expression in germ cells lacking NELF.

| S-EPMC10197597 | biostudies-literature

RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis [RNA-Seq]

Project description:Male germ cell development requires precise regulation of gene activity in a cell-type and stage-specific manner, with perturbations in gene expression during spermatogenesis associated with infertility. Here, we use steady-state, nascent and single-cell RNA sequencing strategies to comprehensively characterize gene expression across male germ cell populations, to dissect the mechanisms of gene control and provide new insights towards therapy. We discover a requirement for pausing of RNA Polymerase II (Pol II) at the earliest stages of sperm differentiation to establish the landscape of gene activity across development. Accordingly, genetic knockout of the Pol II pause-inducing factor NELF in immature germ cells blocks differentiation to spermatids. Further, we uncover unanticipated roles for Pol II pausing in the regulation of meiosis during spermatogenesis, with the presence of paused Pol II associated with double strand break (DSB) formation, and disruption of meiotic gene expression and DSB repair in germ cells lacking NELF.

2023-12-18 | GSE228453 | GEO

RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis[PRO-Seq]

Project description:Male germ cell development requires precise regulation of gene activity in a cell-type and stage-specific manner, with perturbations in gene expression during spermatogenesis associated with infertility. Here, we use steady-state, nascent and single-cell RNA sequencing strategies to comprehensively characterize gene expression across male germ cell populations, to dissect the mechanisms of gene control and provide new insights towards therapy. We discover a requirement for pausing of RNA Polymerase II (Pol II) at the earliest stages of sperm differentiation to establish the landscape of gene activity across development. Accordingly, genetic knockout of the Pol II pause-inducing factor NELF in immature germ cells blocks differentiation to spermatids. Further, we uncover unanticipated roles for Pol II pausing in the regulation of meiosis during spermatogenesis, with the presence of paused Pol II associated with double strand break (DSB) formation, and disruption of meiotic gene expression and DSB repair in germ cells lacking NELF.

2023-12-18 | GSE228452 | GEO

RNA polymerase II pausing is essential for appropriate gene expression and meiosis during spermatogenesis

Project description:Male germ cell development requires precise regulation of gene activity, in a cell-type and stage-specific manner, with perturbations in gene expression during spermatogenesis associated with infertility. Here, we use both steady-state and nascent RNA sequencing strategies to comprehensively characterize gene expression across male germ cell populations, to dissect the mechanisms of gene control and provide new insights towards therapy. We discover a requirement for pausing of RNA Polymerase II (Pol II) at the earliest stages of sperm differentiation to establish the landscape of gene activity across development. Accordingly, genetic knockout of the Pol II pause-inducing factor NELF in immature germ cells impairs differentiation to mature spermatids. Moreover, our work reveals unanticipated roles for Pol II pausing in the regulation of meiosis during spermatogenesis, with the presence of paused Pol II influencing double strand break formation by Spo11, and disruption of Spo11 expression in germ cells lacking NELF.

2023-12-18 | GSE228722 | GEO

RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis

Project description:RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis

| PRJNA949943 | ENA

RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis [RNA-Seq]

Project description:RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis [RNA-Seq]

| PRJNA949951 | ENA

RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis[PRO-Seq]

Project description:RNA polymerase II pausing is essential during spermatogenesis for appropriate gene expression and completion of meiosis[PRO-Seq]

| PRJNA949942 | ENA

RNA polymerase II pausing is essential for appropriate gene expression and meiosis during spermatogenesis

Project description:RNA polymerase II pausing is essential for appropriate gene expression and meiosis during spermatogenesis

| PRJNA951360 | ENA

The m6A reader PRRC2A is essential for meiosis I completion during spermatogenesis.

Project description:N6-methyladenosine (m6A) and its reader proteins YTHDC1, YTHDC2, and YTHDF2 have been shown to exert essential functions during spermatogenesis. However, much remains unknown about m6A regulation mechanisms and the functions of specific readers during the meiotic cell cycle. Here, we show that the m6A reader Proline rich coiled-coil 2A (PRRC2A) is essential for male fertility. Germ cell-specific knockout of Prrc2a causes XY asynapsis and impaired meiotic sex chromosome inactivation in late-prophase spermatocytes. Moreover, PRRC2A-null spermatocytes exhibit delayed metaphase entry, chromosome misalignment, and spindle disorganization at metaphase I and are finally arrested at this stage. Sequencing data reveal that PRRC2A decreases the RNA abundance or improves the translation efficiency of targeting transcripts. Specifically, PRRC2A recognizes spermatogonia-specific transcripts and downregulates their RNA abundance to maintain the spermatocyte expression pattern during the meiosis prophase. For genes involved in meiotic cell division, PRRC2A improves the translation efficiency of their transcripts. Further, co-immunoprecipitation data show that PRRC2A interacts with several proteins regulating mRNA metabolism or translation (YBX1, YBX2, PABPC1, FXR1, and EIF4G3). Our study reveals post-transcriptional functions of PRRC2A and demonstrates its critical role in the completion of meiosis I in spermatogenesis.

| S-EPMC10039029 | biostudies-literature

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