PROJECT D PRDM9 is an important regulator of recombination and transcription;it is uniquely expressed during meiosis;and its absence arrests meiotic progress and gametogenesis. Thus Prdm9 has evolved functionally to serve specific meiotic and gametogenic requirements;however, the full range and nature of these requirements is not known. In particular, the roles of PRDMS in the sequence of meiotic events, its pattern of expression, its relationship to meiotically relevant nuclear structures and domains, and how the structure of PRDMS mediates its varied enzymatic effects are all unknown. The broad, long-term goals of this project are to resolve these issues by using both cellular and genetic strategies to identify mechanisms of when (Aim 1), where (Aim 2) and how (Aim 3) PRDMS promotes recombination site activation, and successful meiosis and spermatogenesis. The results of Aim 1 will establish which steps in meiosis are subject to PRDMS control and whether PRDMS function is required continuously or only at specific meiotic or spermatogenic substages. Because preliminary evidence suggests that PRDMS occupancy in nuclei may be transient, this is important for determining when chromatin marks are set, when recombination site selection occurs, and when transcriptional control is possible. The results of Aim 2 will determine localization of the PRDMS protein and use both PrdmQ mutants and well established mouse meiosis mutant models to determine the extent to which PRDMS function is required for or dependent on proteins crucial for meiosis, including those mediating chromosome synapsis and recombination. The results of Aim 3 will determine which functions of PRDMS in meiosis and gametogenesis require either its histone methyltransferase activity or its transcription-regulating KRAB domain. Recent results suggest that non-coding RNAs are among the transcriptional targets of PRDMS. In cooperation with goals of Projects B and C, this aim will define PRDMS-specific genomic sequence targets and functional clusters of sequences epigenetically marked and/or with transcription regulated by specific PRDMS domains and their relationship to PRDMS-dependent recombination hotspots. Together, the results of this project will complement the other Projects by setting PRDMS function into the well-defined sequence of meiotic chromosome events, more specifically resolving the molecular basis for its multiple roles, and contributing to understanding how PRDMS fits into the larger network of proteins that control meiosis and gametogenesis. An important facet of PRDMS biology addressed by this project is that PRDMS is a major determinant of fertility;the aberrant recombination and failed meiosis that occur in its absence can lead to sterility or birth defects in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM099640-02
Application #
8723852
Study Section
Special Emphasis Panel (ZRG1-GGG-F)
Project Start
Project End
Budget Start
2014-08-01
Budget End
2015-07-31
Support Year
2
Fiscal Year
2014
Total Cost
$314,652
Indirect Cost
$134,851
Name
Jackson Laboratory
Department
Type
DUNS #
042140483
City
Bar Harbor
State
ME
Country
United States
Zip Code
04609
Narasimhan, Vagheesh M; Hunt, Karen A; Mason, Dan et al. (2016) Health and population effects of rare gene knockouts in adult humans with related parents. Science 352:474-7
Huang, Fang; Sirinakis, George; Allgeyer, Edward S et al. (2016) Ultra-High Resolution 3D Imaging of Whole Cells. Cell 166:1028-40
Ball, Robyn L; Fujiwara, Yasuhiro; Sun, Fengyun et al. (2016) Regulatory complexity revealed by integrated cytological and RNA-seq analyses of meiotic substages in mouse spermatocytes. BMC Genomics 17:628
Sun, Fengyun; Fujiwara, Yasuhiro; Reinholdt, Laura G et al. (2015) Nuclear localization of PRDM9 and its role in meiotic chromatin modifications and homologous synapsis. Chromosoma 124:397-415
Didion, John P; Morgan, Andrew P; Clayshulte, Amelia M-F et al. (2015) A multi-megabase copy number gain causes maternal transmission ratio distortion on mouse chromosome 2. PLoS Genet 11:e1004850
Baker, Christopher L; Petkova, Pavlina; Walker, Michael et al. (2015) Multimer Formation Explains Allelic Suppression of PRDM9 Recombination Hotspots. PLoS Genet 11:e1005512
Baker, Christopher L; Kajita, Shimpei; Walker, Michael et al. (2015) PRDM9 drives evolutionary erosion of hotspots in Mus musculus through haplotype-specific initiation of meiotic recombination. PLoS Genet 11:e1004916
Walker, Michael; Billings, Timothy; Baker, Christopher L et al. (2015) Affinity-seq detects genome-wide PRDM9 binding sites and reveals the impact of prior chromatin modifications on mammalian recombination hotspot usage. Epigenetics Chromatin 8:31
Baker, Christopher L; Walker, Michael; Kajita, Shimpei et al. (2014) PRDM9 binding organizes hotspot nucleosomes and limits Holliday junction migration. Genome Res 24:724-32