| 2002 |
Ctr9 is a component of the Paf1/RNA polymerase II complex (Paf1C), distinct from the Srb-mediator form of Pol II holoenzyme. Tandem affinity purification and mass spectrometry established that Ctr9 associates with Paf1, Cdc73, Leo1, Rtf1, and Pol II. Genetic epistasis showed deletion of PAF1 or CTR9 produces similar severe pleiotropic phenotypes that are not additive when combined, placing them in the same pathway. |
Tandem affinity purification, mass spectrometry, genetic epistasis (double-deletion analysis) |
Molecular and cellular biology |
High |
11884586
|
| 1996 |
CTR9/p150TSP is a nuclear phosphoprotein containing multiple tetratricopeptide repeat (TPR) domains that binds specifically to SH2 domains. The TPR module mediates homotypic protein-protein interactions in transfected cells. The C-terminal serine/glutamic acid-rich region is essential for SH2 binding, which depends on serine/threonine phosphorylation but not tyrosine phosphorylation. |
Biochemical purification from B cell lysates, cDNA cloning, transfection with deletion mutants, phosphorylation-dependent binding assays |
The Journal of biological chemistry |
Medium |
8636124
|
| 2018 |
Crystal structures of both human and yeast Ctr9/Paf1 subcomplexes reveal that they assemble into heterodimers with similar conformations via an interface between the TPR module of Ctr9 and Paf1. Formation of the Ctr9/Paf1 heterodimer is required for assembly of the full yeast Paf1C and for yeast viability. Disruption of this interface greatly reduces histone H3 methylation in vivo. |
Crystal structure determination (X-ray crystallography), interface mutagenesis, yeast viability assay, histone modification analysis in vivo |
Nature communications |
High |
30228257
|
| 2013 |
CTR9 (as part of PAFc) constitutively associates with the 5'-coding region of the c-Fos locus and controls the elongation block by regulating NELF and SPT5 chromatin association. CTR9 depletion increases serine 5- and serine 2-phosphorylated Pol II at the unstimulated c-Fos locus, increases CDK9 association, reduces NELF binding, and enhances SPT5 recruitment. IL-6-induced JAK2 kinase activity controls CTR9 chromatin dissociation at this locus. |
ChIP assay, siRNA knockdown, kinase inhibitor (AG-490) treatment, analysis of Pol II phosphorylation states |
PloS one |
Medium |
23593388
|
| 2013 |
Ctr9 knockdown in mouse preimplantation embryos causes loss of histone H3K36me3, failure to correctly specify ICM/TE lineages at the blastocyst stage, and aberrant expression of imprinted genes without altering allele-specific DNA methylation. These phenotypes are similar to those produced by knockdown of Setd2 (the H3K36me3 writer) or Rtf1. |
siRNA knockdown in mouse embryos, immunofluorescence for H3K36me3, allele-specific expression analysis, genetic epistasis with Setd2 and Rtf1 knockdown |
Developmental biology |
Medium |
24036311
|
| 2015 |
Ctr9 enhances ERα protein stability, promotes recruitment of ERα and RNAPII to estrogen-regulated loci, and stimulates transcription elongation and transcription-coupled histone modifications in ERα+ breast cancer cells. Knockdown of Ctr9 (but not other PAFc subunits) nearly completely erases estrogen-regulated transcriptional response and alters cell morphology, proliferative capacity, and tamoxifen sensitivity. |
siRNA knockdown, ChIP, co-immunoprecipitation, ERα stability assay, transcriptome analysis, cell proliferation and morphology assays |
Genes & development |
Medium |
26494790
|
| 2016 |
Loss of Ctr9 in the Drosophila nervous system (null mutant) reduces global H3K4me3 levels, increases neuroblast numbers and nervous system proliferation, and downregulates neuropeptide genes; it also upregulates E2f1 and alters Notch pathway target gene expression. A human CTR9 cDNA transgene rescues lethality of the Drosophila Ctr9 null mutant, demonstrating functional conservation. |
Drosophila null mutation, human cDNA rescue transgene, immunostaining for H3K4me3, genome-wide transcriptome analysis, clonal analysis |
G3 (Bethesda, Md.) |
Medium |
27520958
|
| 2016 |
Drosophila CG2469 encodes a functional Ctr9 homolog; both human and Drosophila Ctr9 localize to nuclei and are enriched in histone locus bodies. Null mutation of Drosophila Ctr9 causes lethality and reduces global H3K4me3 in ovary clones. A human CTR9 cDNA transgene rescues the lethality, confirming functional conservation. |
Sequence analysis, nuclear localization by immunofluorescence, null mutation analysis, H3K4me3 immunostaining in clones, human cDNA rescue |
G3 (Bethesda, Md.) |
Medium |
27678520
|
| 2015 |
Ctr9 physically interacts with the dopamine transporter (DAT) via residues YKF in the first half of the DAT C-terminus, as demonstrated by yeast two-hybrid, GST pulldown, and co-immunoprecipitation. In mammalian cells, Ctr9 partially colocalizes with DAT at the plasma membrane and dramatically enhances DAT-mediated dopamine uptake by increasing the number of DAT transporters at the plasma membrane. Deletion mutagenesis demonstrated that the SH2 domain of Ctr9 is required for its nuclear localization. |
Yeast two-hybrid, GST pulldown, co-immunoprecipitation, co-localization imaging, dopamine uptake assay, deletion mutagenesis |
The Journal of biological chemistry |
Medium |
26048990
|
| 2014 |
CTR9 occupies the coding region of the Il17a gene in naive T cells and dissociates under Th17-polarizing conditions; its depletion induces IL-17 expression and enhances Th17 differentiation. IL-6 directly represses CTR9 promoter activity, creating a feed-forward loop. Lentiviral CTR9 overexpression in joints of collagen-induced arthritis mice reduced arthritis severity and CD4+IL-17+ T cell frequency. |
ChIP assay, siRNA knockdown, promoter reporter assay, lentiviral overexpression in vivo (mouse arthritis model), flow cytometry |
Journal of immunology |
Medium |
24420920
|
| 2018 |
In yeast, Paf1 and Ctr9 (core PAF1C subunits) specifically maintain low levels of telomere repeat-containing RNA (TERRA), while Cdc73, Leo1, and Rtf1 have lesser effects. Double-mutant analysis showed Paf1 and Ctr9 can regulate TERRA independently of Sir4, Rat1, and Trf4 (previously known TERRA regulators), and the data suggest they do so by affecting both transcription and degradation of TERRA. |
Northern blot/TERRA quantification in deletion mutants, genetic epistasis (double mutants with sir4Δ, rat1Δ, trf4Δ) |
Nucleic acids research |
Medium |
29145644
|
| 2022 |
CTR9 controls genome-wide H3K27me3 levels by regulating PRC2 subtype equilibrium. Loss of CTR9 leads to global expansion of H3K27me3, increased PRC2 chromatin recruitment, and a switch from the less active PRC2.2 to the more active PRC2.1 subtype. These effects are reversed by CTR9 restoration, and CTR9 depletion renders breast cancer cells hypersensitive to PRC2 inhibitors. |
Inducible and stable CTR9 knockdown, quantitative histone modification profiling, ChIP-seq for H3K27me3 and PRC2 subunits, PRC2 subtype biochemical analysis, cell viability assays with PRC2 inhibitors |
Nucleic acids research |
Medium |
35137163
|
| 2022 |
CTR9 counteracts EZH2-mediated H3K27me3 deposition in human mesenchymal stem cells. CTR9 knockdown causes gain of H3K27me3 and blocks osteoblast/chondrocyte differentiation; this block is partially rescued by EZH2 inhibitors. CTR9 regulates BMP-2 secretion and membrane anchorage, and the BMP-SMAD pathway is impaired by CTR9 knockdown but rescued by BMP-2 supplementation. |
siRNA knockdown, EZH2 inhibitor rescue, ChIP for H3K27me3, BMP-2 supplementation rescue, in vivo ectopic osteogenesis assay, transcriptome analysis |
Science advances |
Medium |
36383652
|
| 2021 |
CTR9 promotes transcription of the oncogene PEG10 via its promoter region in hepatocellular carcinoma cells. CTR9 knockdown reduces PEG10 expression, increases p21 and p27, and decreases MMP2/MMP9, while overexpression has the opposite effects. These oncogenic roles were confirmed in a xenograft mouse model. |
siRNA knockdown, CTR9 overexpression, promoter reporter assay, western blot, xenograft mouse model |
Acta pharmacologica Sinica |
Low |
34876700
|
| 2023 |
SIAH1 physically interacts with CTR9 (identified by yeast two-hybrid and confirmed by co-immunoprecipitation) and ubiquitinates CTR9 via K48-linked polyubiquitination, leading to proteasomal degradation of CTR9 in HCC cells. SIAH1 inhibits EMT of HCC cells through negative regulation of CTR9 protein levels. |
Yeast two-hybrid, co-immunoprecipitation, ubiquitination assay (K48-linkage specificity), proteasome inhibitor assay, EMT functional assays |
Carcinogenesis |
Medium |
37038329
|
| 2016 |
Genome-wide ChIP-seq demonstrated that Ctr9 knockdown dramatically decreases global chromatin occupancy of ERα and RNAPII in response to estrogen in ERα+ breast cancer cells, establishing that Ctr9 controls ERα-target gene expression by regulating global ERα and RNAPII chromatin binding, independently of other PAFc subunits. |
ChIP-seq for ERα and RNAPII, inducible Ctr9 knockdown, parallel knockdown of other PAFc subunits |
BMC genomics |
Medium |
27829357
|
| 2022 |
De novo CTR9 missense variants (p.Glu15Asp, p.Pro25Arg) associated with neurodevelopmental disorder show stronger affinity to PAF1 protein in pull-down assays compared to wild-type CTR9. In zebrafish, ctr9 knockout causes motor defects and telencephalon enlargement; introduction of human CTR9 mutants failed to rescue these phenotypes, and mutant mRNA overexpression caused telencephalon enlargement, indicating dominant-negative activity. |
Pull-down assay (mutant vs wild-type binding to PAF1), zebrafish ctr9 knockout, human CTR9 mRNA rescue experiment, overexpression of mutant mRNA in zebrafish |
Human molecular genetics |
Medium |
35717577
|
| 2012 |
Proteome analysis of a ctr9Δ yeast strain by 2D gel electrophoresis revealed proteome changes not fully explained by Paf1C functions, and Ctr9 has been described as a triple-helical DNA binding protein, suggesting functions independent of the Paf1 complex. |
2D gel electrophoresis proteomics of ctr9Δ yeast strain |
Biochimica et biophysica acta |
Low |
22446411
|