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A role for alternative splicing in circadian control of exocytosis and glucose homeostasis.

Genes & development · 2020 · Vol. 34 (15-16) · pp. 1089-1105

Abstract

The circadian clock is encoded by a negative transcriptional feedback loop that coordinates physiology and behavior through molecular programs that remain incompletely understood. Here, we reveal rhythmic genome-wide alternative splicing (AS) of pre-mRNAs encoding regulators of peptidergic secretion within pancreatic β cells that are perturbed in <i>Clock</i><sup>-/-</sup> and <i>Bmal1</i><sup>-/-</sup> β-cell lines. We show that the RNA-binding protein THRAP3 (thyroid hormone receptor-associated protein 3) regulates circadian clock-dependent AS by binding to exons at coding sequences flanking exons that are more frequently skipped in clock mutant β cells, including transcripts encoding <i>Cask</i> (<i>calcium/calmodulin-dependent serine protein kinase</i>) and <i>Madd</i> (<i>MAP kinase-activating death domain</i>). Depletion of THRAP3 restores expression of the long isoforms of <i>Cask</i> and <i>Madd</i>, and mimicking exon skipping in these transcripts through antisense oligonucleotide delivery in wild-type islets reduces glucose-stimulated insulin secretion. Finally, we identify shared networks of alternatively spliced exocytic genes from islets of rodent models of diet-induced obesity that significantly overlap with clock mutants. Our results establish a role for pre-mRNA alternative splicing in β-cell function across the sleep/wake cycle.

Publication Types

["Journal Article", "Research Support, N.I.H., Extramural", "Research Support, Non-U.S. Gov't", "Research Support, U.S. Gov't, Non-P.H.S."]

Keywords

MeSH Terms

["ARNTL Transcription Factors", "Alternative Splicing", "Animals", "CLOCK Proteins", "Cells, Cultured", "Circadian Clocks", "Death Domain Receptor Signaling Adaptor Proteins", "Exocytosis", "Glucose", "Guanine Nucleotide Exchange Factors", "Guanylate Kinases", "Homeostasis", "Insulin Secretion", "Insulin-Secreting Cells", "Islets of Langerhans", "Male", "Mice, Inbred C57BL", "Nuclear Proteins", "Obesity", "Synaptosomal-Associated Protein 25", "Transcription Factors"]

Funding

R01 HG004659 NHGRI NIH HHS (United States)
F32 HL143978 NHLBI NIH HHS (United States)
T32 GM145427 NIGMS NIH HHS (United States)
T32 GM008666 NIGMS NIH HHS (United States)
R01 DK090625 NIDDK NIH HHS (United States)
CIHR (Canada)
R01 HL061322 NHLBI NIH HHS (United States)
U54 AR052646 NIAMS NIH HHS (United States)
R01 DK113011 NIDDK NIH HHS (United States)
T32 GM008152 NIGMS NIH HHS (United States)
R01 NS047726 NINDS NIH HHS (United States)
R01 DK050203 NIDDK NIH HHS (United States)
R01 AG065988 NIA NIH HHS (United States)
T32 GM144295 NIGMS NIH HHS (United States)
R01 DK100814 NIDDK NIH HHS (United States)
T32 DK007169 NIDDK NIH HHS (United States)
T32 HL007909 NHLBI NIH HHS (United States)
F30 DK116481 NIDDK NIH HHS (United States)
R01 HL140938 NHLBI NIH HHS (United States)
P30 DK020595 NIDDK NIH HHS (United States)
P60 DK020595 NIDDK NIH HHS (United States)
P01 AG011412 NIA NIH HHS (United States)

Linked Datasets (1)

GSE146916 GSE via ncbi_elink
GEO

A role for alternative splicing in circadian control of insulin secretion and glucose homeostasis

Mus musculus
23 data files
FileTypeSize
BKO_RNA_1_R1.fastq RNA-Seq 1.4 GB
BKO_RNA_2_R1.fastq RNA-Seq 1.8 GB
BKO_RNA_3_R1.fastq RNA-Seq 1.2 GB
BKO_THRAP3_ChIPseq_Input.fq.gz ChIP-Seq 476.9 MB
BKO_THRAP3_ChIPseq_rep1.fq.gz ChIP-Seq 580.2 MB
BKO_THRAP3_ChIPseq_rep2.fq.gz ChIP-Seq 650.8 MB
BKO_THRAP3_eCLIP_input1.fastq.gz OTHER 1.3 GB
BKO_THRAP3_eCLIP_input2.fastq.gz OTHER 471.0 MB
BKO_THRAP3_eCLIP_IP1.fastq.gz OTHER 907.5 MB
BKO_THRAP3_eCLIP_IP2.fastq.gz OTHER 685.2 MB
CKO_RNA_1_R1.fastq RNA-Seq 1.3 GB
CKO_RNA_2_R1.fastq RNA-Seq 1.1 GB
CKO_RNA_3_R1.fastq RNA-Seq 1.1 GB
WT_RNA_1_R1.fastq RNA-Seq 1.4 GB
WT_RNA_2_R1.fastq RNA-Seq 1.3 GB
WT_RNA_3_R1.fastq RNA-Seq 1.1 GB
WT_THRAP3_ChIPseq_Input.fq.gz ChIP-Seq 513.4 MB
WT_THRAP3_ChIPseq_rep1.fq.gz ChIP-Seq 567.5 MB
WT_THRAP3_ChIPseq_rep2.fq.gz ChIP-Seq 578.3 MB
WT_THRAP3_eCLIP_input1.fastq.gz OTHER 1.5 GB
WT_THRAP3_eCLIP_input2.fastq.gz OTHER 1.1 GB
WT_THRAP3_eCLIP_IP1.fastq.gz OTHER 1.7 GB
WT_THRAP3_eCLIP_IP2.fastq.gz OTHER 930.9 MB

Potentially Related Datasets (3)

These accessions were text-mined from the PMC full text. They may be referenced for comparison, cited from other studies, or otherwise mentioned without being primary data for this paper.

GSE69889 GSE GEO
GEO

Genome-wide Circadian Control of Transcription at Active Enhancers Regulates Insulin Secretion and Diabetes Risk

GSE92602 GSE GEO
GEO

Identification of islet-enriched long non-coding RNAs contributing to beta-cell failure in type 2 diabetes

GSE95156 GSE GEO
GEO

Pancreatic alpha and beta-cellular clocks have distinct molecular properties and impact on islet hormone secretion and gene expression

Analysis Pipelines (4)

eCLIP geo_data_processing GSE146916
RNA-seq geo_data_processing GSE69889
RNA-seq geo_data_processing GSE92602
geo_data_processing GSE95156