Next Generation Sequencing Analysis Identifies Genes Whose Expression is Regulated by SIRT1 and IGF2BP2
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA758066 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15622866.fastq.gz | Fastqsanger.gz | |||
| SRR15622867.fastq.gz | Fastqsanger.gz | |||
| SRR15622868.fastq.gz | Fastqsanger.gz | |||
| SRR15622869.fastq.gz | Fastqsanger.gz | |||
| SRR15622870.fastq.gz | Fastqsanger.gz |
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Project description:This study shows how cells that are depleted of sirtuin1 (SIRT1) regulate IGF2BP2 to promote the degradation of the transcript that encodes ATP6V1A, a major catalytic subunit of the v-ATPase. IGF2BP2 and SIRT1 actions lead to altered gene expression by determining epigenetic modifications and by modulating RNA stability. This raises the question of whether the regulation of IGF2BP2 impacts additional transcripts, similarly to what we found for the transcript encoding ATP6V1A. The purpose of this RNA sequencing analysis is to identify genes whose expression is regulated similarly to ATP6V1A.
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Project description:Insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) binds various RNA transcripts and functions as a tumor promoter, although little is known regarding the mechanisms that regulate its roles in RNA metabolism (1-3). Here we find that IGF2BP2 binds to the 3’ untranslated region of the transcript encoding ATP6V1A, a major catalytic subunit of the vacuolar ATPase (v-ATPase), and regulates its degradation in a lysine acetylation and SIRT1-dependent manner. We show that the regulation of IGF2BP2 is significantly compromised in breast cancer cells where SIRT1 expression is low or knocked-down. Reduced SIRT1 expression leads to an increase in acetylated IGF2BP2, which enables IGF2BP2 to recruit the XRN2 nuclease to the ATP6V1A transcript, resulting in its degradation and a reduction in the expression of functional v-ATPase complexes. This impairs lysosomal activity and produces a cellular secretome consisting of increased numbers of exosomes enriched in ubiquitinated protein cargo and soluble hydrolases including cathepsins, which combine to promote tumor progression and invasiveness. Collectively, these findings describe a previously unrecognized role for IGF2BP2 in mediating the degradation of an mRNA transcript essential for lysosomal function and highlight how its sirtuin-regulated acetylation state can have significant biological and disease consequences.
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Project description:Insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) binds various RNA transcripts and functions as a tumor promoter, although little is known regarding the mechanisms that regulate its roles in RNA metabolism (1-3). Here we find that IGF2BP2 binds to the 3’ untranslated region of the transcript encoding ATP6V1A, a major catalytic subunit of the vacuolar ATPase (v-ATPase), and regulates its degradation in a lysine acetylation and SIRT1-dependent manner. We show that the regulation of IGF2BP2 is significantly compromised in breast cancer cells where SIRT1 expression is low or knocked-down. Reduced SIRT1 expression leads to an increase in acetylated IGF2BP2, which enables IGF2BP2 to recruit the XRN2 nuclease to the ATP6V1A transcript, resulting in its degradation and a reduction in the expression of functional v-ATPase complexes. This impairs lysosomal activity and produces a cellular secretome consisting of increased numbers of exosomes enriched in ubiquitinated protein cargo and soluble hydrolases including cathepsins, which combine to promote tumor progression and invasiveness. Collectively, these findings describe a previously unrecognized role for IGF2BP2 in mediating the degradation of an mRNA transcript essential for lysosomal function and highlight how its sirtuin-regulated acetylation state can have significant biological and disease consequences.
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Project description:The expression of IGF2BP2 in GCs from PCOS patients was detected using RT-qPCR and western blot. We captured IGF2BP2-interacting transcripts, global transcriptome together with alternative splicing by RNA immunoprecipitation sequencing (RIP-seq) and RNA sequencing (RNA-seq). KGN cells transfected with IGF2BP2 overexpressing plasmids and nuclear factor 1 C-type (NFIC) siRNAs, were applied to CCK-8, EdU and TUNEL assays. IGF2BP2 was highly expressed in GCs from PCOS patients. As a RBP, it preferentially bound to the 3’and 5’UTRs of mRNAs with GGAC motif and a newly found GAAG motif. The overexpression of IGF2BP2 changed the transcriptome profile of KGN cells. IGF2BP2 functioned to regulate alternative splicing events (ASEs) and promote cell proliferation through inhibiting exon skipping events of NFIC. In conclusion, we demonstrated that IGF2BP2 promotes granulosa cell proliferation via regulating alternative splicing of NFIC in PCOS. The findings help to better understand the roles of IGF2BP2 in the pathogenesis of PCOS.
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Project description:Sirt1, a NAD-dependent deacetylase, has been shown involved in many complex biological functions. We previously demonstrated that Sirt-1 gain of function is neuroprotective in a model of neurodegenenration. We recently found that Sirt1 loss of function (Sirt1Δ mouse) impairs learning, memory, and synaptic plasticity. We used microarray screening to identify potential miRNAs that may be regulated by Sirt1 and ivolved in learning and memory. Total RNA from the hippocampus of SIRT1Δ and control mice was extracted and used for microRNA expression level evaluation.
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