# RNA Pol II Pausing is Critical for Spermatogenesis and Male Fertility

> **NIH NIH R01** · UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN · 2021 · $327,599

## Abstract

PROJECT SUMMARY
Successful completion of spermatogenesis relies upon precise spatiotemporal expression of distinct subsets of
differentiation markers within the seminiferous epithelium. Failure to express genes at the correct time leads to
arrested spermatogenesis and male infertility. The transcriptional mechanisms regulating this process, however,
are not well understood. Our work has established that RNA Pol II pausing is critical for maintaining precise
spatiotemporal gene expression during spermatogenesis. Paused RNA Pol II at the promoter ensures precise
and rapid onset of gene transcription. This mechanism is particularly relevant to spermatogenesis wherein
synchronous transcription of cohorts of genes is critical for morphogenesis and differentiation. The central
hypothesis of this proposal is that RNA Pol II pausing occurs genome-wide in germ cells and is critical for the
success of spermatogenesis. We have identified the TAR DNA binding protein of 43 kD (TDP-43) as a key player
in maintaining paused Pol II at a target gene promoter in male germ cells. A corollary is that TDP-43 regulates
pausing of a subset of genes in germ cells. TDP-43 is evolutionarily conserved and expressed in the mouse and
human testis. We have found that TDP-43 is essential for spermatogenesis; conditional knockout of TDP-43 in
germ cells led to maturation arrest and male infertility. Specific Aim 1 will test the overarching hypothesis that
Pol II pausing is a key regulator of spatiotemporal gene transcription by determining the genome-wide occupancy
of Pol II pause machinery in germ cells and seek biochemical validation for pausing. Aim 2 will study the
mechanism of Pol II pausing in germ cells; test the hypothesis that TDP-43 recruits the pause machinery to a
subset of promoters, map its interactions with pause factors and determine functional significance. Specific Aim
3 will test the hypothesis that loss of TDP-43 disrupts Pol II pausing at TDP-43 target promoters in germ cells
leading to male infertility. In human cells including embryonic stem cells, Pol II pausing has been shown to play
a pivotal role in regulation of gene transcription. The present study is significant because for the first time it will
expand knowledge on a mechanism regulating the male germ cell transcriptome. The outcome will have a major
impact on the understanding of the genetic basis of idiopathic male infertility. Thus, the proposed studies are
aligned with high priority topic areas identified by the Fertility and Infertility (FI) Branch of NICHD: Genetic basis
of idiopathic infertility and Models for infertility.

## Key facts

- **NIH application ID:** 10199764
- **Project number:** 5R01HD094546-04
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
- **Principal Investigator:** PRABHAKARA P REDDI
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $327,599
- **Award type:** 5
- **Project period:** 2018-08-21 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10199764

## Citation

> US National Institutes of Health, RePORTER application 10199764, RNA Pol II Pausing is Critical for Spermatogenesis and Male Fertility (5R01HD094546-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10199764. Licensed CC0.

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