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The non-radioactive SUnSET assay to measure glucose-induced protein synthesis in human β-cells

ABSTRACT: Glucose regulates the expression of a limited number of genes in human pancreatic β-cells at the transcriptional level. Here, we asked whether glucose intervenes at post-transcriptional steps and regulates protein translation in human β-cell lines. First, we showed the involvement of the m-TOR pathway in glucose-related signaling. We next used the Surface SEnsing of Translation (SUnSET) technique that is based on puromycin incorporation into newly-translated proteins to show that glucose treatment increased protein translation. Among the list of glucose-induced proteins, we selected the proconvertase PCSK1 as a promising target whose translation was induced within minutes following glucose treatment. We finally performed global proteomic analysis by mass spectrometry to characterize newly-translated proteins upon glucose treatment. We identified many proteins involved in translation, glycolysis, TCA metabolism and insulin secretion. Taken together, our study demonstrates that while glucose poorly modulates gene expression in human β-cells, it plays major role at the translational level.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Islet Of Langerhans, Pancreatic Beta Cell Line

DISEASE(S): Type 2 Diabetes Mellitus

SUBMITTER: Chiara guerrera

LAB HEAD: Chiara Guerrera

PROVIDER: PXD023996 | Pride | 2021-07-27

REPOSITORIES: Pride

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