AGO4 Regulates Entry into Meiosis and Influences Silencing of Sex Chromosomes in the Male Mouse Germline

Modzelewski, A; Holmes, Rebecca; Hilz, Stephanie; Grimson, Andrew; Cohen, Paula E.

doi:10.1016/j.devcel.2012.07.003

Cited by 90 publications

(102 citation statements)

References 83 publications

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“…We find that Ago2 is the most prominent Ago protein in all tissues analyzed, followed by Ago1 and Ago3. Ago4 is found mainly in testes, which is consistent with the testes phenotype of Ago4 knockout mice (23) (Fig. 2E).…”

Section: Specific Isolation and Characterization Of Ago Protein Complsupporting

confidence: 69%

Biochemical isolation of Argonaute protein complexes by Ago-APP

Hauptmann

Schraivogel

Bruckmann

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

121

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During microRNA (miRNA)-guided gene silencing, Argonaute (Ago) proteins interact with a member of the TNRC6/GW protein family. Here we used a short GW protein-derived peptide fused to GST and demonstrate that it binds to Ago proteins with high affinity. This allows for the simultaneous isolation of all Ago protein complexes expressed in diverse species to identify associated proteins, small RNAs, or target mRNAs. We refer to our method as “Ago protein Affinity Purification by Peptides“ (Ago-APP). Furthermore, expression of this peptide competes for endogenous TNRC6 proteins, leading to global inhibition of miRNA function in mammalian cells.

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Section: Specific Isolation and Characterization Of Ago Protein Complsupporting

confidence: 69%

Biochemical isolation of Argonaute protein complexes by Ago-APP

Hauptmann

Schraivogel

Bruckmann

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

121

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“…However, based on the finding that most micro(mi)RNAs on the mouse X are highly expressed when all X-linked protein coding genes are silenced by MSCI (Song et al 2009), Yan and proposed that these MSCI-escaping miRNAs play a key role in regulating X inactivation in the male germ line. Preliminary support for this hypothesis is provided by an association between failed MSCI and disproportionate downregulation of miRNAs on the X vs. the autosomes in male lab mice lacking AGO4, a key protein in the miRNA pathway (Modzelewski et al 2012).The expression patterns in the seven X introgression F 1 's assayed in this study demonstrate a strong association between X overexpression and an interval on the proximal half Figure 3 Hybrid males with more X overexpression have worse reproductive phenotypes. Significant negative correlations between X overexpression (mean DDC T ) and relative testis weight (RTW, R 2 = 0.33, P = 0.003), sperm count (R 2 = 0.34, P = 0.003), and the percentage of normal sperm (R 2 = 0.71, P , 0.0001), measured in the 24 F 1 males used in the whole-testis expression study (see Figure 2A for genotypes).…”

mentioning

confidence: 54%

“…However, based on the finding that most micro(mi)RNAs on the mouse X are highly expressed when all X-linked protein coding genes are silenced by MSCI (Song et al 2009), Yan and McCarrey (2009) proposed that these MSCI-escaping miRNAs play a key role in regulating X inactivation in the male germ line. Preliminary support for this hypothesis is provided by an association between failed MSCI and disproportionate downregulation of miRNAs on the X vs. the autosomes in male lab mice lacking AGO4, a key protein in the miRNA pathway (Modzelewski et al 2012).…”

Section: Disruption Of Mscimentioning

confidence: 82%

“…Burgoyne and colleagues proposed that MSCI fails under these conditions because this key protein is unavailable Burgoyne et al 2009). A recent report of excess asynapsis in musculus PWK · C57BL/6J pachytene spermatocytes (Modzelewski et al 2012) indicates that this phenotype may exist in the musculus PWK · domesticus LEWES cross, a hypothesis that we plan to evaluate by quantifying asynapsis, DSBs, and BRCA1 distribution in pachytene chromosome spreads. …”

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confidence: 99%

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Meiotic Sex Chromosome Inactivation Is Disrupted in Sterile Hybrid Male House Mice

2013

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In male mammals, the X and Y chromosomes are transcriptionally silenced in primary spermatocytes by meiotic sex chromosome inactivation (MSCI) and remain repressed for the duration of spermatogenesis. Here, we test the longstanding hypothesis that disrupted MSCI might contribute to the preferential sterility of heterogametic hybrid males. We studied a cross between wildderived inbred strains of Mus musculus musculus and M. m. domesticus in which sterility is asymmetric: F 1 males with a M. m. musculus mother are sterile or nearly so while F 1 males with a M. m. domesticus mother are normal. In previous work, we discovered widespread overexpression of X-linked genes in the testes of sterile but not fertile F 1 males. Here, we ask whether this overexpression is specifically a result of disrupted MSCI. To do this, we isolated cells from different stages of spermatogenesis and measured the expression of several genes using quantitative PCR. We found that X overexpression in sterile F 1 primary spermatocytes is coincident with the onset of MSCI and persists in postmeiotic spermatids. Using a series of recombinant X genotypes, we then asked whether X overexpression in hybrids is controlled by cis-acting loci across the X chromosome. We found that it is not. Instead, one large interval in the proximal portion of the M. m. musculus X chromosome is associated with both overexpression and the severity of sterility phenotypes in hybrids. These results demonstrate a strong association between X-linked hybrid male sterility and disruption of MSCI and suggest that transacting loci on the X are important for the transcriptional regulation of the X chromosome during spermatogenesis. FORTY years ago, Lifschytz and Lindsley (1972) proposed the provocative hypothesis that the X chromosome is inactivated during male meiosis and that disruption of this process could lead to sterility. The idea that failure of meiotic sex chromosome inactivation (MSCI) might cause sterility is significant in speciation genetics since it provides a possible explanation for the widespread observation that in crosses between species, sterility typically appears first in the heterogametic sex (Haldane 1922;Forejt 1996;Presgraves 2008).Whether MSCI takes place in Drosophila has been debated for several decades (Kremer et al. 1986;McKee and Handel 1993;Hense et al. 2007;Vibranovski et al. 2009). Current evidence suggests that expression on the X chromosome is suppressed relative to the autosomes during spermatogenesis but that this suppression precedes meiosis and thus does not reflect MSCI (Meiklejohn et al. 2011; but see Vibranovski et al. 2012). Likewise, the sex chromosomes are not differentially silenced in chicken oocytes (Guioli et al. 2012). Therefore, failed MSCI cannot explain hybrid sterility in all female-heterogametic taxa. In therian mammals, however, MSCI is well established (Solari 1974;McKee and Handel 1993;Namekawa et al. 2007). Although no MSCI-essential loci have been found on the sex chromosomes, many of the autosomal genes t...

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“…The presence and functions of sncRNAs in mammalian spermatogenesis have been analyzed in different studies (Papaioannou and Nef 2010;McIver et al 2012;Modzelewski et al 2012;Yadav and Kotaja 2014). In particular, the miRNAs have been defined as robust regulators of germ cell development (Kedde and Agami 2008).…”

Section: Introductionmentioning

confidence: 99%

Diversity and functional convergence of small noncoding RNAs in male germ cell differentiation and fertilization

García-López

Alonso

Cárdenas

et al. 2015

RNA

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The small noncoding RNAs (sncRNAs) are considered as post-transcriptional key regulators of male germ cell development. In addition to microRNAs (miRNAs) and PIWI-interacting RNAs (piRNAs), other sncRNAs generated from small nucleolar RNAs (snoRNAs), tRNAs, or rRNAs processing may also play important regulatory roles in spermatogenesis. By next-generation sequencing (NGS), we characterized the sncRNA populations detected at three milestone stages in male germ differentiation: primordial germ cells (PGCs), pubertal spermatogonia cells, and mature spermatozoa. To assess their potential transmission through the spermatozoa during fertilization, the sncRNAs of mouse oocytes and zygotes were also analyzed. Both, microRNAs and snoRNA-derived small RNAs are abundantly expressed in PGCs but transiently replaced by piRNAs in spermatozoa and endo-siRNAs in oocytes and zygotes. Exhaustive analysis of miRNA sequence variants also shows an increment of noncanonical microRNA forms along male germ cell differentiation. RNAs-derived from tRNAs and rRNAs interacting with PIWI proteins are not generated by the ping-pong pathway and could be a source of primary piRNAs. Moreover, our results strongly suggest that the small RNAs-derived from tRNAs and rRNAs are interacting with PIWI proteins, and specifically with MILI. Finally, computational analysis revealed their potential involvement in post-transcriptional regulation of mRNA transcripts suggesting functional convergence among different small RNA classes in germ cells and zygotes.

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AGO4 Regulates Entry into Meiosis and Influences Silencing of Sex Chromosomes in the Male Mouse Germline

Cited by 90 publications

References 83 publications

Biochemical isolation of Argonaute protein complexes by Ago-APP

Biochemical isolation of Argonaute protein complexes by Ago-APP

Meiotic Sex Chromosome Inactivation Is Disrupted in Sterile Hybrid Male House Mice

Diversity and functional convergence of small noncoding RNAs in male germ cell differentiation and fertilization

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