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TAF4b Regulates Oocyte-Specific Genes Essential for Meiosis.

Grive KJ, Gustafson EA, Seymour KA, Baddoo M, Schorl C, Golnoski K, Rajkovic A, Brodsky AS, Freiman RN - PLoS Genet. (2016)

Bottom Line: To further explore the potential mechanism of TAF4b in promoting ovarian follicle development, we analyzed global gene expression at multiple time points in the human fetal ovary.This computational analysis revealed coordinate expression of human TAF4B and critical regulators and effectors of meiosis I including SYCP3, YBX2, STAG3, and DAZL.Accordingly, TAF4b occupies the proximal promoters of many essential meiosis and oogenesis regulators, including Stra8, Dazl, Figla, and Nobox, and is required for their proper expression.

View Article: PubMed Central - PubMed

Affiliation: MCB Graduate Program, Brown University, Providence, Rhode Island, United States of America.

ABSTRACT
TAF4b is a gonadal-enriched subunit of the general transcription factor TFIID that is implicated in promoting healthy ovarian aging and female fertility in mice and humans. To further explore the potential mechanism of TAF4b in promoting ovarian follicle development, we analyzed global gene expression at multiple time points in the human fetal ovary. This computational analysis revealed coordinate expression of human TAF4B and critical regulators and effectors of meiosis I including SYCP3, YBX2, STAG3, and DAZL. To address the functional relevance of this analysis, we turned to the embryonic Taf4b-deficient mouse ovary where, for the first time, we demonstrate, severe deficits in prophase I progression as well as asynapsis in Taf4b-deficient oocytes. Accordingly, TAF4b occupies the proximal promoters of many essential meiosis and oogenesis regulators, including Stra8, Dazl, Figla, and Nobox, and is required for their proper expression. These data reveal a novel TAF4b function in regulating a meiotic gene expression program in early mouse oogenesis, and support the existence of a highly conserved TAF4b-dependent gene regulatory network promoting early oocyte development in both mice and women.

No MeSH data available.


Related in: MedlinePlus

TAF4b targets the promoters of critical meiosis and oogenesis regulators.(A) Wild-type E18.5 ovarian chromatin pulled down by antibodies against TAF4b or IgG and then qPCR-amplified using primers against the proximal promoters of Stra8, Dazl, Figlα, Nobox and a non-genic region upstream of Nobox. qPCR demonstrates enriched recovery of these proximal promoters with TAF4b-precipitated chromatin relative to IgG. (B) The Nobox proximal promoter, in particular, demonstrates enrichment for TAF4b, in contrast to a non-genic region upstream of the promoter. For all analyses, data from each primer set were normalized to the E18.5 mouse ovary genomic DNA input levels and represented as a percentage of that DNA input. Each qPCR reaction was performed in triplicate and averaged. Error bars indicate the normalized standard deviation resulting from experimental triplicate qPCR reactions.
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pgen.1006128.g005: TAF4b targets the promoters of critical meiosis and oogenesis regulators.(A) Wild-type E18.5 ovarian chromatin pulled down by antibodies against TAF4b or IgG and then qPCR-amplified using primers against the proximal promoters of Stra8, Dazl, Figlα, Nobox and a non-genic region upstream of Nobox. qPCR demonstrates enriched recovery of these proximal promoters with TAF4b-precipitated chromatin relative to IgG. (B) The Nobox proximal promoter, in particular, demonstrates enrichment for TAF4b, in contrast to a non-genic region upstream of the promoter. For all analyses, data from each primer set were normalized to the E18.5 mouse ovary genomic DNA input levels and represented as a percentage of that DNA input. Each qPCR reaction was performed in triplicate and averaged. Error bars indicate the normalized standard deviation resulting from experimental triplicate qPCR reactions.

Mentions: While TAF4b is evidently required for proper regulation of meiosis I progression, the nature of this regulation was unclear. To test if TAF4b directly occupies the proximal promoters of essential meiosis genes, we performed ChIP from wild-type E18.5 ovaries using a validated anti-TAF4b antibody (S2 Fig) or a negative control IgG antibody, and performed quantitative PCR. Strikingly, immunoprecipitated chromatin bound by TAF4b included the proximal promoters of Stra8 and Dazl (Fig 5A). As oogenesis regulators Figlα and Nobox are known downstream targets of DAZL [35], these promoters were also tested and found to be directly bound by TAF4b (Fig 5A and 5B). TAF4b occupancy at these important loci is specific, as genomic regions not expected to be occupied by TAF4b, including a non-genic region 50kb upstream of Nobox were not enriched for TAF4b (Fig 5B). Quantitative PCR results were validated by gel electrophoresis and visualization of amplified proximal promoters (S3 Fig).


TAF4b Regulates Oocyte-Specific Genes Essential for Meiosis.

Grive KJ, Gustafson EA, Seymour KA, Baddoo M, Schorl C, Golnoski K, Rajkovic A, Brodsky AS, Freiman RN - PLoS Genet. (2016)

TAF4b targets the promoters of critical meiosis and oogenesis regulators.(A) Wild-type E18.5 ovarian chromatin pulled down by antibodies against TAF4b or IgG and then qPCR-amplified using primers against the proximal promoters of Stra8, Dazl, Figlα, Nobox and a non-genic region upstream of Nobox. qPCR demonstrates enriched recovery of these proximal promoters with TAF4b-precipitated chromatin relative to IgG. (B) The Nobox proximal promoter, in particular, demonstrates enrichment for TAF4b, in contrast to a non-genic region upstream of the promoter. For all analyses, data from each primer set were normalized to the E18.5 mouse ovary genomic DNA input levels and represented as a percentage of that DNA input. Each qPCR reaction was performed in triplicate and averaged. Error bars indicate the normalized standard deviation resulting from experimental triplicate qPCR reactions.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4920394&req=5

pgen.1006128.g005: TAF4b targets the promoters of critical meiosis and oogenesis regulators.(A) Wild-type E18.5 ovarian chromatin pulled down by antibodies against TAF4b or IgG and then qPCR-amplified using primers against the proximal promoters of Stra8, Dazl, Figlα, Nobox and a non-genic region upstream of Nobox. qPCR demonstrates enriched recovery of these proximal promoters with TAF4b-precipitated chromatin relative to IgG. (B) The Nobox proximal promoter, in particular, demonstrates enrichment for TAF4b, in contrast to a non-genic region upstream of the promoter. For all analyses, data from each primer set were normalized to the E18.5 mouse ovary genomic DNA input levels and represented as a percentage of that DNA input. Each qPCR reaction was performed in triplicate and averaged. Error bars indicate the normalized standard deviation resulting from experimental triplicate qPCR reactions.
Mentions: While TAF4b is evidently required for proper regulation of meiosis I progression, the nature of this regulation was unclear. To test if TAF4b directly occupies the proximal promoters of essential meiosis genes, we performed ChIP from wild-type E18.5 ovaries using a validated anti-TAF4b antibody (S2 Fig) or a negative control IgG antibody, and performed quantitative PCR. Strikingly, immunoprecipitated chromatin bound by TAF4b included the proximal promoters of Stra8 and Dazl (Fig 5A). As oogenesis regulators Figlα and Nobox are known downstream targets of DAZL [35], these promoters were also tested and found to be directly bound by TAF4b (Fig 5A and 5B). TAF4b occupancy at these important loci is specific, as genomic regions not expected to be occupied by TAF4b, including a non-genic region 50kb upstream of Nobox were not enriched for TAF4b (Fig 5B). Quantitative PCR results were validated by gel electrophoresis and visualization of amplified proximal promoters (S3 Fig).

Bottom Line: To further explore the potential mechanism of TAF4b in promoting ovarian follicle development, we analyzed global gene expression at multiple time points in the human fetal ovary.This computational analysis revealed coordinate expression of human TAF4B and critical regulators and effectors of meiosis I including SYCP3, YBX2, STAG3, and DAZL.Accordingly, TAF4b occupies the proximal promoters of many essential meiosis and oogenesis regulators, including Stra8, Dazl, Figla, and Nobox, and is required for their proper expression.

View Article: PubMed Central - PubMed

Affiliation: MCB Graduate Program, Brown University, Providence, Rhode Island, United States of America.

ABSTRACT
TAF4b is a gonadal-enriched subunit of the general transcription factor TFIID that is implicated in promoting healthy ovarian aging and female fertility in mice and humans. To further explore the potential mechanism of TAF4b in promoting ovarian follicle development, we analyzed global gene expression at multiple time points in the human fetal ovary. This computational analysis revealed coordinate expression of human TAF4B and critical regulators and effectors of meiosis I including SYCP3, YBX2, STAG3, and DAZL. To address the functional relevance of this analysis, we turned to the embryonic Taf4b-deficient mouse ovary where, for the first time, we demonstrate, severe deficits in prophase I progression as well as asynapsis in Taf4b-deficient oocytes. Accordingly, TAF4b occupies the proximal promoters of many essential meiosis and oogenesis regulators, including Stra8, Dazl, Figla, and Nobox, and is required for their proper expression. These data reveal a novel TAF4b function in regulating a meiotic gene expression program in early mouse oogenesis, and support the existence of a highly conserved TAF4b-dependent gene regulatory network promoting early oocyte development in both mice and women.

No MeSH data available.


Related in: MedlinePlus