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A miR-130a-YAP positive feedback loop promotes organ size and tumorigenesis.

Shen S, Guo X, Yan H, Lu Y, Ji X, Li L, Liang T, Zhou D, Feng XH, Zhao JC, Yu J, Gong XG, Zhang L, Zhao B - Cell Res. (2015)

Bottom Line: Organ size determination is one of the most intriguing unsolved mysteries in biology.Here we report that the YAP signaling is sustained through a novel microRNA-dependent positive feedback loop. miR-130a, which is directly induced by YAP, could effectively repress VGLL4, an inhibitor of YAP activity, thereby amplifying the YAP signals.Furthermore, the Drosophila Hippo pathway target bantam functionally mimics miR-130a by repressing the VGLL4 homolog SdBP/Tgi.

View Article: PubMed Central - PubMed

Affiliation: Life Sciences Institute and Innovation Center for Cell Signaling Network Hangzhou, Zhejiang 310058, China.

ABSTRACT
Organ size determination is one of the most intriguing unsolved mysteries in biology. Aberrant activation of the major effector and transcription co-activator YAP in the Hippo pathway causes drastic organ enlargement in development and underlies tumorigenesis in many human cancers. However, how robust YAP activation is achieved during organ size control remains elusive. Here we report that the YAP signaling is sustained through a novel microRNA-dependent positive feedback loop. miR-130a, which is directly induced by YAP, could effectively repress VGLL4, an inhibitor of YAP activity, thereby amplifying the YAP signals. Inhibition of miR-130a reversed liver size enlargement induced by Hippo pathway inactivation and blocked YAP-induced tumorigenesis. Furthermore, the Drosophila Hippo pathway target bantam functionally mimics miR-130a by repressing the VGLL4 homolog SdBP/Tgi. These findings reveal an evolutionarily conserved positive feedback mechanism underlying robustness of the Hippo pathway in size control and tumorigenesis.

No MeSH data available.


Related in: MedlinePlus

miR-130a is a direct target gene of YAP-TEAD. (A) YAP induces miR-130a expression in an S94-dependent manner. pri-miR-130a expression levels in indicated MCF10A stable cells were determined by quantitative RT-PCR. Experiments were performed in triplicates. (B) Knockdown of TEADs reduces miR-130a level. Cells were infected with the shRNA targeting TEAD1/3/4. The levels of mature miR-130a were determined. Experiments were performed in triplicates. (C) YAP represses miR-130a sensor activity in an S94-dependent manner. HepG2 cells were transfected with miR-130a sensor and YAP WT or mutants for luciferase assay. Experiments were performed in duplicates. (D)miR-130a genomic structure. miR-130a promoter is in blue, pri-miR-130a encoding region is in green, and pre-miR-130a encoding region is in red. Regions amplified by ChIP-PCR primers are labeled as P1 to P11. Regions amplified by primers used in RT-PCR confirmation of TSS are labeled as a-e. The proximal promoter cloned for luciferase reporter is indicated. (E, F) YAP and TEAD bind to the proximal promoter of miR-130a. HepG2 cells expressing Flag-YAP-5SA (E) or native HepG2 cells (F) were processed for ChIP with control IgG or antibodies against TEAD1 and YAP followed by quantitative PCR. Experiments were performed in duplicates. (G) YAP activates miR-130a promoter reporter in a TEAD-dependent manner. WT or mutant miR-130a reporters were transfected with or without YAP and TEAD2 into HEK293T cells for luciferase assay. Experiments were performed in duplicates.
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fig2: miR-130a is a direct target gene of YAP-TEAD. (A) YAP induces miR-130a expression in an S94-dependent manner. pri-miR-130a expression levels in indicated MCF10A stable cells were determined by quantitative RT-PCR. Experiments were performed in triplicates. (B) Knockdown of TEADs reduces miR-130a level. Cells were infected with the shRNA targeting TEAD1/3/4. The levels of mature miR-130a were determined. Experiments were performed in triplicates. (C) YAP represses miR-130a sensor activity in an S94-dependent manner. HepG2 cells were transfected with miR-130a sensor and YAP WT or mutants for luciferase assay. Experiments were performed in duplicates. (D)miR-130a genomic structure. miR-130a promoter is in blue, pri-miR-130a encoding region is in green, and pre-miR-130a encoding region is in red. Regions amplified by ChIP-PCR primers are labeled as P1 to P11. Regions amplified by primers used in RT-PCR confirmation of TSS are labeled as a-e. The proximal promoter cloned for luciferase reporter is indicated. (E, F) YAP and TEAD bind to the proximal promoter of miR-130a. HepG2 cells expressing Flag-YAP-5SA (E) or native HepG2 cells (F) were processed for ChIP with control IgG or antibodies against TEAD1 and YAP followed by quantitative PCR. Experiments were performed in duplicates. (G) YAP activates miR-130a promoter reporter in a TEAD-dependent manner. WT or mutant miR-130a reporters were transfected with or without YAP and TEAD2 into HEK293T cells for luciferase assay. Experiments were performed in duplicates.

Mentions: To determine how YAP induces miR-130a, we checked whether the induction depends on TEADs. Indeed, mutation of YAP serine 94 (S94), a residue critical for TEAD interaction15, abrogated miR-130a induction (Figure 2A). Furthermore, knockdown of TEADs strongly reduced miR-130a expression (Figure 2B and Supplementary information, Figure S2A, S2B). To confirm the regulation of miR-130a activity by YAP-TEAD, we constructed a miR-130a sensor fusing a stretch of four perfect miR-130a binding sites 3′ to the luciferase gene. Function of the sensor was confirmed by co-transfection with miR-130a mimic and inhibitor (Supplementary information, Figure S2C). Indeed, YAP but not the S94A mutant clearly repressed miR-130a sensor activity (Figure 2C). Thus miR-130a is a target of YAP-TEAD transcriptional complex.


A miR-130a-YAP positive feedback loop promotes organ size and tumorigenesis.

Shen S, Guo X, Yan H, Lu Y, Ji X, Li L, Liang T, Zhou D, Feng XH, Zhao JC, Yu J, Gong XG, Zhang L, Zhao B - Cell Res. (2015)

miR-130a is a direct target gene of YAP-TEAD. (A) YAP induces miR-130a expression in an S94-dependent manner. pri-miR-130a expression levels in indicated MCF10A stable cells were determined by quantitative RT-PCR. Experiments were performed in triplicates. (B) Knockdown of TEADs reduces miR-130a level. Cells were infected with the shRNA targeting TEAD1/3/4. The levels of mature miR-130a were determined. Experiments were performed in triplicates. (C) YAP represses miR-130a sensor activity in an S94-dependent manner. HepG2 cells were transfected with miR-130a sensor and YAP WT or mutants for luciferase assay. Experiments were performed in duplicates. (D)miR-130a genomic structure. miR-130a promoter is in blue, pri-miR-130a encoding region is in green, and pre-miR-130a encoding region is in red. Regions amplified by ChIP-PCR primers are labeled as P1 to P11. Regions amplified by primers used in RT-PCR confirmation of TSS are labeled as a-e. The proximal promoter cloned for luciferase reporter is indicated. (E, F) YAP and TEAD bind to the proximal promoter of miR-130a. HepG2 cells expressing Flag-YAP-5SA (E) or native HepG2 cells (F) were processed for ChIP with control IgG or antibodies against TEAD1 and YAP followed by quantitative PCR. Experiments were performed in duplicates. (G) YAP activates miR-130a promoter reporter in a TEAD-dependent manner. WT or mutant miR-130a reporters were transfected with or without YAP and TEAD2 into HEK293T cells for luciferase assay. Experiments were performed in duplicates.
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fig2: miR-130a is a direct target gene of YAP-TEAD. (A) YAP induces miR-130a expression in an S94-dependent manner. pri-miR-130a expression levels in indicated MCF10A stable cells were determined by quantitative RT-PCR. Experiments were performed in triplicates. (B) Knockdown of TEADs reduces miR-130a level. Cells were infected with the shRNA targeting TEAD1/3/4. The levels of mature miR-130a were determined. Experiments were performed in triplicates. (C) YAP represses miR-130a sensor activity in an S94-dependent manner. HepG2 cells were transfected with miR-130a sensor and YAP WT or mutants for luciferase assay. Experiments were performed in duplicates. (D)miR-130a genomic structure. miR-130a promoter is in blue, pri-miR-130a encoding region is in green, and pre-miR-130a encoding region is in red. Regions amplified by ChIP-PCR primers are labeled as P1 to P11. Regions amplified by primers used in RT-PCR confirmation of TSS are labeled as a-e. The proximal promoter cloned for luciferase reporter is indicated. (E, F) YAP and TEAD bind to the proximal promoter of miR-130a. HepG2 cells expressing Flag-YAP-5SA (E) or native HepG2 cells (F) were processed for ChIP with control IgG or antibodies against TEAD1 and YAP followed by quantitative PCR. Experiments were performed in duplicates. (G) YAP activates miR-130a promoter reporter in a TEAD-dependent manner. WT or mutant miR-130a reporters were transfected with or without YAP and TEAD2 into HEK293T cells for luciferase assay. Experiments were performed in duplicates.
Mentions: To determine how YAP induces miR-130a, we checked whether the induction depends on TEADs. Indeed, mutation of YAP serine 94 (S94), a residue critical for TEAD interaction15, abrogated miR-130a induction (Figure 2A). Furthermore, knockdown of TEADs strongly reduced miR-130a expression (Figure 2B and Supplementary information, Figure S2A, S2B). To confirm the regulation of miR-130a activity by YAP-TEAD, we constructed a miR-130a sensor fusing a stretch of four perfect miR-130a binding sites 3′ to the luciferase gene. Function of the sensor was confirmed by co-transfection with miR-130a mimic and inhibitor (Supplementary information, Figure S2C). Indeed, YAP but not the S94A mutant clearly repressed miR-130a sensor activity (Figure 2C). Thus miR-130a is a target of YAP-TEAD transcriptional complex.

Bottom Line: Organ size determination is one of the most intriguing unsolved mysteries in biology.Here we report that the YAP signaling is sustained through a novel microRNA-dependent positive feedback loop. miR-130a, which is directly induced by YAP, could effectively repress VGLL4, an inhibitor of YAP activity, thereby amplifying the YAP signals.Furthermore, the Drosophila Hippo pathway target bantam functionally mimics miR-130a by repressing the VGLL4 homolog SdBP/Tgi.

View Article: PubMed Central - PubMed

Affiliation: Life Sciences Institute and Innovation Center for Cell Signaling Network Hangzhou, Zhejiang 310058, China.

ABSTRACT
Organ size determination is one of the most intriguing unsolved mysteries in biology. Aberrant activation of the major effector and transcription co-activator YAP in the Hippo pathway causes drastic organ enlargement in development and underlies tumorigenesis in many human cancers. However, how robust YAP activation is achieved during organ size control remains elusive. Here we report that the YAP signaling is sustained through a novel microRNA-dependent positive feedback loop. miR-130a, which is directly induced by YAP, could effectively repress VGLL4, an inhibitor of YAP activity, thereby amplifying the YAP signals. Inhibition of miR-130a reversed liver size enlargement induced by Hippo pathway inactivation and blocked YAP-induced tumorigenesis. Furthermore, the Drosophila Hippo pathway target bantam functionally mimics miR-130a by repressing the VGLL4 homolog SdBP/Tgi. These findings reveal an evolutionarily conserved positive feedback mechanism underlying robustness of the Hippo pathway in size control and tumorigenesis.

No MeSH data available.


Related in: MedlinePlus