Transcriptomics

Dataset Information

3

Studying the effects of the TEN1-ICD on transcriptional regulation


ABSTRACT: Teneurins are large type II transmembrane proteins that are necessary for the normal development of the central nervous system (CNS). While many studies highlight the significance of teneurins, especially during development, there is only limited information known about the molecular mechanisms of function. Previous studies have shown that the N-terminal intracellular domain (ICD) of teneurins can be cleaved at the membrane and subsequently translocates to the nucleus where it can influence gene transcription. Target genes as well as mechanisms have yet to be elucidated, and thus we are investigating the transcriptional activity of the human teneurin-1 ICD in this study. For the whole transcriptome analysis of TEN1-ICD overexpression, we used a modified and improved tet-system. Two separate vectors are required to make the cell line stable. One contains the tet-activating domain fused to a glucocorticoid binding domain (GBD). The other contains the tetO operator sequences directly upstream of a CMV promoter and the gene to be overexpressed. BS149 cells were first transfected with pirtetR-GBD and made stable by Puromycin selection, and then after further transfection with either ptetO-eGFP-His (negative control) or ptetO-TEN1-ICD-eGFP-His by Hygromycin selection. The stable BS149 cell lines were split into three 10 cm Petri dishes each. The triplicate cell lines were cultured once before induction with Dexamethasone and Doxycycline. The overexpressing cells were then FACS-sorted directly into RLT lysis buffer (Qiagen) at a 3:1 volume ratio of lysis buffer to cells in PBS, 24 h post-induction.

REANALYSED by: E-GEOD-61704

OTHER RELATED OMICS DATASETS IN: E-GEOD-61705

ORGANISM(S): Homo sapiens  

SUBMITTER: Jonas Schöler   Ruth Chiquet-Ehrismann  Tim Roloff  Jacqueline Ferralli  Stéphane Thiry 

PROVIDER: E-GEOD-61704 | ArrayExpress | 2015-02-04

SECONDARY ACCESSION(S): GSE61704PRJNA261901

REPOSITORIES: GEO, ArrayExpress

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Publications

The intracellular domain of teneurin-1 induces the activity of microphthalmia-associated transcription factor (MITF) by binding to transcriptional repressor HINT1.

Schöler Jonas J   Ferralli Jacqueline J   Thiry Stéphane S   Chiquet-Ehrismann Ruth R  

The Journal of biological chemistry 20150203 13


Teneurins are large type II transmembrane proteins that are necessary for the normal development of the CNS. Although many studies highlight the significance of teneurins, especially during development, there is only limited information known about the molecular mechanisms of function. Previous studies have shown that the N-terminal intracellular domain (ICD) of teneurins can be cleaved at the membrane and subsequently translocates to the nucleus, where it can influence gene transcription. Becau  ...[more]

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