<i>Drosophila CG2469</i> Encodes a Homolog of Human CTR9 and Is Essential for Development

Chaturvedi, Dhananjay; Inaba, Mayu; Scoggin, Shane

doi:10.1534/g3.116.035196

Cited by 17 publications

(21 citation statements)

References 27 publications

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“…1E), suggesting they act downstream of PIWI-initiated silencing in fly ovaries. Since ATU is the homolog of Leo1, a component of the yeast PAF1 complex, we tested other Drosophila PAF1 complex factors, such as PAF1, RTF1, CTR9 (also called CG2469, [28, 29]), and HYRAX (HYR, homolog of CDC73 [30]). Only HYR knockdown impaired PIWI-directed silencing, while CTR9 knockdown lacked impact on the Flam reporters (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Drosophila PAF1 Modulates PIWI/piRNA Silencing Capacity

et al. 2017

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SUMMARY To test the directness of factors in initiating PIWI-directed gene silencing, we employed a Piwi-interacting RNA (piRNA)-targeted reporter assay in Drosophila OSS cells [1]. This assay confirmed direct silencing roles for piRNA biogenesis factors and PIWI-associated factors [2–12], but suggested that chromatin-modifying proteins may act downstream of the initial silencing event. Our data also revealed that RNA Polymerase II associated proteins like PAF1 and RTF1 antagonizes PIWI-directed silencing. PAF1 knockdown enhances PIWI silencing of reporters when piRNAs target the transcript region proximal to the promoter. Loss of PAF1 suppresses endogenous transposable element (TE) transcript maturation, whereas a subset of gene transcripts and long-non-coding RNAs adjacent to TE insertions are affected by PAF1 knockdown in a similar fashion to piRNA-targeted reporters. Additionally, transcription activation at specific TEs and TE-adjacent loci during PIWI knockdown is suppressed when PIWI and PAF1 levels are both reduced. Our study suggests a mechanistic conservation between fission yeast PAF1 repressing AGO1/siRNA-directed silencing [13, 14] and Drosophila PAF1 opposing PIWI/piRNA-directed silencing.

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Section: Resultsmentioning

confidence: 99%

“…Homozygous mutants and whole-fly knockdowns of Drosophila PAF1 factors are lethal [28–30, 32]. To test the impact of Drosophila PAF1 in ovary development, we used transgenic RNAi to knock down dPaf1 and dRtf1 with germline Nos-Gal4 and soma Tj-Gal4 drivers (Figure S2).…”

Section: Resultsmentioning

confidence: 99%

Drosophila PAF1 Modulates PIWI/piRNA Silencing Capacity

et al. 2017

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“…yeast) and multicellular organisms when CTR9 is depleted. While Ctr9 is not an essential gene in S. cerevisiae (Chen et al, 2002; Massoni-Laporte et al, 2012), CTR9 knockout causes early embryonic lethality in higher eukaryotic organisms such as Drosophila (Chaturvedi et al, 2016), zebrafish (Akanuma et al, 2007) and mouse (Zhang et al, 2013). The requirement of CTR9 in preimplantation development of mice resembles the phenotypes that result from lacking core PRC2 subunits SUZ12, EZH2 and EED (Pasini et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

Transcriptional elongation machinery controls vulnerability of breast cancer cells to PRC2 inhibitors

Huang

et al. 2020

Preprint

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1 2 CTR9 is the scaffold subunit in Paf1c, a multifunctional complex regulating multiple steps 3 of RNA Pol II-mediated transcription. Using inducible and stable CTR9 knockdown breast 4 cancer cell lines, we discovered that the expression of a subset of KDMs, including 5 KDM6A and Jarid2, is strictly controlled by CTR9. Global analyses of histone modifications 6 revealed a significant increase of H3K27me3 upon loss of CTR9. Loss of CTR9 results in 7 a decrease of H3K4me3 and H3K36me3 in gene bodies, and elevated levels and genome-8 wide expansion of H3K27me3. Mechanistically, CTR9 depletion triggers a PRC2 subtype 9 switching from PRC2.2 to PRC2.1. As a consequence, CTR9 depletion generates 10 vulnerability that renders breast cancer cells hypersensitive to PRC2 inhibitors. Our 11 findings that CTR9 demarcates PRC2-mediated H3K27me3 levels and genomic 12 distribution, provide a unique mechanism of transition from transcriptionally active to 13 repressive chromatin states and sheds light on the biological functions of CTR9 in 14 development and cancer. 15 16 17 18 19 20 21 22 23 4 RESULTS 1 2 CTR9 regulates a subset of histone lysine demethylases (KDMs) in ER+ breast 3 cancer cells 4 Data mining of the CTR9-regulated transcriptome in CTR9 inducible knockdown (KD) 5 MCF7 cells (MCF7-tet-on-shCtr9) (Figure 1A) (Zeng and Xu, 2016) identified a subset of 6 KDM genes that mirrors the expression of CTR9, including KDM1A, KDM2B, KDM3B, 7KDM5B, KDM6A and JARID2. We found that these KDM genes were down-regulated 8 when CTR9 was knocked down by shRNA in MCF7 cells after a 7-day doxycycline (Dox) 9 treatment ( Figure 1B) regardless of whether 17β-estradiol (E2) was present or absent. 10RNAPII binding peaks at the transcription start site (TSS) of several KDM genes also 11 decreased in response to CTR9 depletion ( Figure S1A). We validated the transcriptome 12 array results by RT-qPCR. Figure 1C shows that silencing of CTR9 after a 7-day Dox 13 treatment reduced the total mRNA levels, and reduced the expression of ribosome-14 associated RNAs, which are indicative of actively transcribed mRNAs, of six KDMs. 15However, the mRNA levels of KDM4B and KDM6B did not significantly change. CTR9's 16 regulation of KDMs was also observed at the protein level. As shown in Figure 1D, the 17 protein levels of six KDMs significantly decreased in the total cell lysates and extracted 18 chromatin fractions. Consistent with their mRNA levels in response to CTR9 depletion, 19 KDM4B and KDM6B protein levels also remained unchanged. To exclude the possibility 20 that this observation was specific to MCF7, or due to off-target effects of anti-CTR9 shRNA, 21 we employed MCF7 cells as well as T47D cells, another well-established ERα+ breast 22 34 35

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“…In the female GSCs, it has recently been shown that the global level of H3K4me3 is decreased upon loss-of-function of the Drosophila ortholog of Ctr9, a component of the Paf1 complex normally required for transcriptional initiation and polyadenylation. However, the functional readout of this global H3K4me3 loss remains unclear ( Chaturvedi et al 2016 ).…”

Section: Mechanisms Regulating Gsc Self-renewal Vsmentioning

confidence: 99%

Protecting and Diversifying the Germline

et al. 2018

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Gametogenesis represents the most dramatic cellular differentiation pathways in both female and male flies. At the genome level, meiosis ensures that diploid germ cells become haploid gametes. At the epigenome level, extensive changes are required to turn on and shut off gene expression in a precise spatiotemporally controlled manner. Research applying conventional molecular genetics and cell biology, in combination with rapidly advancing genomic tools have helped us to investigate (1) how germ cells maintain lineage specificity throughout their adult reproductive lifetime; (2) what molecular mechanisms ensure proper oogenesis and spermatogenesis, as well as protect genome integrity of the germline; (3) how signaling pathways contribute to germline-soma communication; and (4) if such communication is important. In this chapter, we highlight recent discoveries that have improved our understanding of these questions. On the other hand, restarting a new life cycle upon fertilization is a unique challenge faced by gametes, raising questions that involve intergenerational and transgenerational epigenetic inheritance. Therefore, we also discuss new developments that link changes during gametogenesis to early embryonic development—a rapidly growing field that promises to bring more understanding to some fundamental questions regarding metazoan development.

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Drosophila CG2469 Encodes a Homolog of Human CTR9 and Is Essential for Development

Cited by 17 publications

References 27 publications

Drosophila PAF1 Modulates PIWI/piRNA Silencing Capacity

Drosophila PAF1 Modulates PIWI/piRNA Silencing Capacity

Transcriptional elongation machinery controls vulnerability of breast cancer cells to PRC2 inhibitors

Protecting and Diversifying the Germline

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