Stable silencing of ZBED6 affects morphology, gene expression and insulin release in insulin-producing islet cells
(English)Manuscript (preprint) (Other academic)
Zbed6 has evolved from a domesticated DNA transposon and encodes a novel transcription factor unique to placental mammals. Here we have investigated the function of ZBED6 in insulin-producing beta cells based on whole transcriptome analysis of MIN6 cells with lentiviral shRNA-mediated stable silencing of either Zbed6 (shZbed6) or mock mRNA (shMock). Zbed6-silencing was associated with altered cell morphology as the shZbed6 cells showed increased neuron-like protrusions compared with shMock cells. ZBED6 appeared as an important transcriptional regulator in islet cells since more than 700 genes showed differential expression in shZbed6 cells when compared with control cells. The most significantly enriched GO categories among differentially expressed genes were neuronal differentiation and cell adhesion, which is consistent with the changes in morphology in the silenced cells. A ChIP-seq analysis identified more than 4,000 putative binding sites in the genome of MIN6 cells and there was a significant overrepresentation of genes with ZBED6 sites among the differentially expressed genes after silencing. This suggests that ZBED6 acts as a transcriptional regulator for many genes in MIN6 cells. The genes showing differential expression included Pdx1, Mafa and Nkx6-1, three crucial transcription factors in beta-cell maturation, which were all up-regulated after Zbed6-silencing. Finally, in shZbed6 MIN6 cells the content and release of insulin was increased. We conclude that ZBED6 is expressed in insulin-producing islet cells and has a significant role for the modulation of cellular functions in this cell type.
Transcriptome analysis, insulin, beta cells
IdentifiersURN: urn:nbn:se:uu:diva-183722OAI: oai:DiVA.org:uu-183722DiVA: diva2:563926