YAP/TAZ enhances P-body formation to promote tumorigenesis
- Department of Colorectal and Anal Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, China
- Shanghai Colorectal Cancer Research Center, China
- State Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, China
Figures
Figure 1 with 1 supplement
YAP/TAZ transcriptionally regulates genes related to P-bodies.
(A) Gene Ontology (GO) analysis of the downregulated genes upon knockdown of YAP/TAZ in HCT116 cells. The graph shows enrichment in the cellular component category. (B) ChIP–qPCR analysis of …
-
Figure 1—source data 1
Original data for the statistical analysis in Figure 1B–D.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
RNA-seq and ChIP-seq analysis of YAP/TEAD’s target genes related to P-bodies.
(A) Heatmap showing the mRNA levels of 76 genes annotated as P-body-related genes, as detected in HCT116 cells by RNA-seq (GSE176475). (B) Representative sequencing TEAD4 ChIP-seq tracks at the SAMD4…
-
Figure 1—figure supplement 1—source data 1
Original data for the heat map in Figure 1—figure supplement 1A.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig1-figsupp1-data1-v1.xlsx
-
Figure 1—figure supplement 1—source data 2
Original file for the Western blot analysis in Figure 1—figure supplement 1C and D.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig1-figsupp1-data2-v1.pdf
Figure 2 with 2 supplements
YAP/TAZ promotes P-body formation in colorectal cancer (CRC) cells.
(A) Immunofluorescence analysis of the P-body markers DDX6 and DCP1A in YAP/TAZ knockdown HCT116 cells. Cells were transfected with YAP/TAZ siRNA for 3 d before processing for immunofluorescence …
-
Figure 2—source data 1
Original data for the statistical analysis in Figure 2A–F.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
YAP/TAZ modulates P-body formation in breast, lung and colorectal cancer cells.
(A) Immunofluorescence analysis of the P-body markers DDX6 and DCP1A in YAP/TAZ knockdown MDA-MB-231 cells. Cells were transfected with YAP/TAZ siRNA for 3 d before processing for immunofluorescence …
-
Figure 2—figure supplement 1—source data 1
Original data for the statistical analysis in Figure 2—figure supplement 1A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig2-figsupp1-data1-v1.xlsx
Figure 2—figure supplement 2
YAP/TAZ modulates P-body formation in untransformed NIH3T3 cells.
(A) Immunofluorescence analysis of DDX6 and LSM14A in NIH3T3 cells expressing YAP5SA and YAP5SA-S94A. Foci were counted in 100 cells per group. (B) Immunofluorescence analysis of DDX6 and LSM14A in …
-
Figure 2—figure supplement 2—source data 1
Original data for the statistical analysis in Figure 2—figure supplement 2A–H.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig2-figsupp2-data1-v1.xlsx
Figure 3 with 1 supplement
SAMD4A, AJUBA, and PNRC1 mediate the regulatory functions of YAP/TAZ in P-body formation.
(A) Immunofluorescence analysis of DDX6 and DCP1A in HCT116 cells stably expressing YAP5SA and YAP5SA-expressing cells transiently transfected with SMAD4A and AJUBA siRNA. Foci were counted in 100 …
-
Figure 3—source data 1
Original data for the statistical analysis in Figure 3A and B.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Knockdown of PNRC1 reverses the attenuated P-body formation induced by YAP/TAZ knockdown.
(A, B) qPCR analysis of the mRNA levels of SAMD4A (A) and AJUBA (B) in HCT116 cells stably expressing FLAG-YAP5SA and YAP5SA and transfected with SMAD4A or AJUBA siRNA. n = 3 biologically …
-
Figure 3—figure supplement 1—source data 1
Original data for the statistical analysis in Figure 3—figure supplement 1A–F.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig3-figsupp1-data1-v1.xlsx
-
Figure 3—figure supplement 1—source data 2
Original file for the western blot analysis in Figure 3—figure supplement 1D.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig3-figsupp1-data2-v1.pdf
Figure 4 with 1 supplement
YAP suppresses PNRC1 gene transcription by recruiting the NuRD complex.
(A) Overexpression of YAP5SA but not YAP5SA-S94A decreased the luciferase activity of the PNRC1 promoter and intron reporters. HEK-293T cells were transfected with the indicated FLAG-YAP5SA and YAP5S…
-
Figure 4—source data 1
Original data for the statistical analysis in Figure 4A–F.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
TEADs and CHD4 mediates YAP-dependent inhibition on PNRC1 gene transcription.
(A) qPCR and western blot analysis of the YAP/TAZ knockdown stable HCT116 cells. (B) ChIP–qPCR analysis of FLAG-YAP5SA, YAP5SA-S94A binding to the PNRC1 promoter and intronic regions in control and …
-
Figure 4—figure supplement 1—source data 1
Original data for the statistical analysis in Figure 4—figure supplement 1A–E.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig4-figsupp1-data1-v1.xlsx
-
Figure 4—figure supplement 1—source data 2
Original file for the western blot analysis in Figure 4—figure supplement 1A.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig4-figsupp1-data2-v1.pdf
Figure 5 with 1 supplement
PNRC1 attenuates the oncogenic function of YAP in colorectal cancer (CRC).
(A) CCK8 proliferation assays of HCT116 cells stably expressing YAP5SA alone or in combination with of PNRC1WT or PNRC1W300A. n = 4 biologically independent samples per group. (B, C) Colony …
-
Figure 5—source data 1
Original data for the statistical analysis in Figure 5A–E.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
PNRC1 is a tumor suppressor gene in CRC.
(A) Downregulated mRNA expression of PNRC1 in colon cancer (COAD) and rectal cancer (READ) colorectal cancer (CRC) datasets in TCGA. The mRNA levels of PNRC1 were extracted from the GEPIA database. …
-
Figure 5—figure supplement 1—source data 1
Original data for the statistical analysis in Figure 5—figure supplement 1B–E.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig5-figsupp1-data1-v1.xlsx
Figure 6 with 4 supplements
Knockdown of P-body-related core genes suppresses the oncogenic function of YAP in colorectal cancer (CRC).
(A) CCK8 proliferation assays of control HCT116 cells with or without knockdown of DCP1A, LSM14A or DDX6 and HCT116 cells stably expressing YAP5SA with or without knockdown of DCP1A, LSM14A, or DDX6.…
-
Figure 6—source data 1
Original data for the statistical analysis in Figure 6A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
Knockdown efficiency and P-body reduction in DCP1A/LSM14A/DDX6 knockdown HCT116 cells.
(A) qPCR and western blot analysis of the knockdown efficiency of DCP1A, LSM14A, and DDX6 in HCT116 cells stably expressing YAP5SA with or without knockdown of DCP1A, LSM14A, or DDX6. n = 3 …
-
Figure 6—figure supplement 1—source data 1
Original data for the statistical analysis in Figure 6—figure supplement 1A and B.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-figsupp1-data1-v1.xlsx
-
Figure 6—figure supplement 1—source data 2
Original file for the western blot analysis in Figure 6—figure supplement 1A.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-figsupp1-data2-v1.pdf
Figure 6—figure supplement 2
Confirmation of DCP1A/ LSM14A/DDX6 knockdown in YAP5SA-expressing HCT116 cells.
(A–C) qPCR and western blot analysis of the knockdown efficiency of DCP1A (A), DDX6 (B), and LSM14A (C) in control HCT116 cells with or without knockdown of DCP1A, LSM14A, or DDX6 and stable YAP5SA …
-
Figure 6—figure supplement 2—source data 1
Original data for the statistical analysis in Figure 6—figure supplement 2A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-figsupp2-data1-v1.xlsx
-
Figure 6—figure supplement 2—source data 2
Original file for the western blot analysis in Figure 6—figure supplement 2A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-figsupp2-data2-v1.pdf
Figure 6—figure supplement 3
Knockdown of DCP1A, LSM14A and DDX6 downregulates cell mitosis and increases cell apoptosis in YAP5SA-expressing HCT116 cells.
(A, B) Cell apoptosis (A) and cell cycle (B) analysis of the control HCT116 cells with or without knockdown of DCP1A, LSM14A, or DDX6 and stable YAP5SA overexpression HCT116 cells with or without …
-
Figure 6—figure supplement 3—source data 1
Original data for the bar plot in Figure 6—figure supplement 3A and B.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-figsupp3-data1-v1.xlsx
Figure 6—figure supplement 4
Knockdown of AJUBA or SAMD4A attenuates the oncogenic function of YAP in HCT116 cells.
(A) Transwell assay of control HCT116 cells with or without knockdown of DDX6 and HCT116 cells stably expressing YAP5SA with or without knockdown of DDX6. n = 3 biologically independent samples per …
-
Figure 6—figure supplement 4—source data 1
Original data for the statistical analysis in Figure 6—figure supplement 4A–D.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-figsupp4-data1-v1.xlsx
-
Figure 6—figure supplement 4—source data 2
Original file for the western blot analysis in Figure 6—figure supplement 4C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig6-figsupp4-data2-v1.pdf
Figure 7 with 5 supplements
DepMap analysis reveals the co-dependencies of YAP/TEAD and P-body core genes in pancancer CRISPR screens.
(A) Positive correlations between the mRNA levels of CTGF and SAMD4A/AJUBA/WTIP in 1393 cancer cell lines. (B) Negative correlation between the mRNA levels of CTGF and PNRC1 in brain cancer cell …
-
Figure 7—source data 1
Original data for the statistical analysis in Figure 7A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-data1-v1.xlsx
Figure 7—figure supplement 1
Correlation analysis of the expression of YAP canonical target genes and YAP-related P-body genes.
(A) Positive correlations between the mRNA levels of YAP target genes (CYR61, AXL, and AMOTL2) and SAMD4A/AJUBA/WTIP in 1393 cancer cell lines. (B) Positive correlations between the mRNA levels of …
-
Figure 7—figure supplement 1—source data 1
Original data for the statistical analysis in Figure 7—figure supplement 1A–D.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp1-data1-v1.xlsx
Figure 7—figure supplement 2
The TEAD1/3 dependency scores are positively correlated with the EDC4/DDX6/LSM14A scores.
Positive correlations between the dependency scores of TEAD1/3 and DDX6/LSM14A/EDC4 in 1070 cancer cell lines. The Chronos dependency scores were extracted from the DepMap database. The negative …
-
Figure 7—figure supplement 2—source data 1
Original data for the statistical analysis in Figure 7—figure supplement 2A.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp2-data1-v1.xlsx
Figure 7—figure supplement 3
Knockdown of DCP1A/LSM14A and overexpression of PNRC1 suppress both cell proliferation and cell migration in A549 lung cancer cells.
(A) qPCR and western blot analysis of the knockdown efficiency of DCP1A / LSM14A or PNRC1 overexpression in A549 cells stably expressing PNRC1WT/PNRC1W300A or with knockdown of DCP1A and LSM14A. (B, …
-
Figure 7—figure supplement 3—source data 1
Original data for the statistical analysis in Figure 7—figure supplement 3A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp3-data1-v1.xlsx
-
Figure 7—figure supplement 3—source data 2
Original file for the western blot analysis in Figure 7—figure supplement 3A.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp3-data2-v1.pdf
Figure 7—figure supplement 4
Knockdown of DCP1A/LSM14A and overexpression of PNRC1 suppress both cell proliferation and cell migration in MCF7 breast cancer cells.
(A) qPCR and western blot analysis of the knockdown efficiency of DCP1A / LSM14A or PNRC1 overexpression in MCF7 cells stably expressing PNRC1WT/PNRC1W300A or with knockdown of DCP1A and LSM14A. (B, …
-
Figure 7—figure supplement 4—source data 1
Original data for the statistical analysis in Figure 7—figure supplement 4A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp4-data1-v1.xlsx
-
Figure 7—figure supplement 4—source data 2
Original file for the western blot analysis in Figure 7—figure supplement 4A.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp4-data2-v1.pdf
Figure 7—figure supplement 5
Knockdown of DCP1A/LSM14A and overexpression of PNRC1 suppress both cell proliferation and cell migration in MDA-MB-231 breast cancer cells.
(A) qPCR and western blot analysis of the knockdown efficiency of DCP1A / LSM14A or PNRC1 overexpression in MDA-MB-231 cells stably expressing PNRC1WT/PNRC1W300A or with knockdown of DCP1A and …
-
Figure 7—figure supplement 5—source data 1
Original data for the statistical analysis in Figure 7—figure supplement 5A–C.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp5-data1-v1.xlsx
-
Figure 7—figure supplement 5—source data 2
Original file for the western blot analysis in Figure 7—figure supplement 5A.
- https://cdn.elifesciences.org/articles/88573/elife-88573-fig7-figsupp5-data2-v1.pdf
Author response image 1
Author response image 2
Author response image 3
Author response image 4
Author response image 5
Author response image 6
Author response image 7
Additional files
-
Supplementary file 1
The expression of P-body genes in YAP/TAZ knockdown HCT116 cells.
The differentially expressed genes in YAP/TAZ knockdown HCT116 cells were analyzed by RNA-seq (GSE176475). The expression levels of genes that were annotated as related to P-bodies are included in this supplementary file.
- https://cdn.elifesciences.org/articles/88573/elife-88573-supp1-v1.xlsx
-
Supplementary file 2
Top 100 correlation for EDC4 in CRISPR (DepMap 22Q1 Public Score, Chronos).
The gene dependency scores of the top 100 genes that are correlated with EDC4 were downloaded from the Cancer Dependency Map (DepMap) database.
- https://cdn.elifesciences.org/articles/88573/elife-88573-supp2-v1.xlsx
-
Supplementary file 3
siRNA, shRNA, and primers used in this study.
The sequences of the siRNA, shRNA, and primers used in this study are included in this supplementary file.
- https://cdn.elifesciences.org/articles/88573/elife-88573-supp3-v1.xlsx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/88573/elife-88573-mdarchecklist1-v1.docx
