Distinct cytoplasmic and nuclear functions of the stress induced protein DDIT3/CHOP/GADD153
Artikel i vetenskaplig tidskrift, 2012

DDIT3, also known as GADD153 or CHOP, encodes a basic leucine zipper transcription factor of the dimer forming C/EBP family. DDIT3 is known as a key regulator of cellular stress response, but its target genes and functions are not well characterized. Here, we applied a genome wide microarray based expression analysis to identify DDIT3 target genes and functions. By analyzing cells carrying tamoxifen inducible DDIT3 expression constructs we show distinct gene expression profiles for cells with cytoplasmic and nuclear localized DDIT3. Of 175 target genes identified only 3 were regulated by DDIT3 in both cellular localizations. More than two thirds of the genes were downregulated, supporting a role for DDIT3 as a dominant negative factor that could act by either cytoplasmic or nuclear sequestration of dimer forming transcription factor partners. Functional annotation of target genes showed cell migration, proliferation and apoptosis/survival as the most affected categories. Cytoplasmic DDIT3 affected more migration associated genes, while nuclear DDIT3 regulated more cell cycle controlling genes. Cell culture experiments confirmed that cytoplasmic DDIT3 inhibited migration, while nuclear DDIT3 caused a G1 cell cycle arrest. Promoters of target genes showed no common sequence motifs, reflecting that DDIT3 forms heterodimers with several alternative transcription factors that bind to different motifs. We conclude that expression of cytoplasmic DDIT3 regulated 94 genes. Nuclear translocation of DDIT3 regulated 81 additional genes linked to functions already affected by cytoplasmic DDIT3. Characterization of DDIT3 regulated functions helps understanding its role in stress response and involvement in cancer and degenerative disorders.

genetics

Promoter Regions

RNA

Tamoxifen

Cytoplasm

genetics

metabolism

genetics

genetics

Transcription Factor CHOP

pharmacology

drug therapy

Gene Expression Profiling

Hormonal

Blotting

Reverse Transcriptase Polymerase Chain Reaction

genetics

Genetic

Biological

pharmacology

Fibroblasts

Cell Movement

metabolism

Tumor Markers

Oligonucleotide Array Sequence Analysis

Messenger

Cell Nucleus

Flow Cytometry

genetics

drug therapy

genetics

Green Fluorescent Proteins

metabolism

drug effects

Cell Proliferation

Antineoplastic Agents

metabolism

Cultured

genetics

Cell Cycle

drug effects

Real-Time Polymerase Chain Reaction

Liposarcoma

Western

Cells

Humans

metabolism

Cell Adhesion

metabolism

metabolism

drug effects

Fibrosarcoma

cytology

genetics

metabolism

Recombinant Proteins

Författare

Alexandra Jauhiainen

Karolinska Institutet

Christer Thomsen

Göteborgs universitet

Linda Strömbom

Göteborgs universitet

Pernilla Grundevik

Göteborgs universitet

Carola Andersson

Göteborgs universitet

Anna Danielsson

Göteborgs universitet

Mattias K Andersson

Göteborgs universitet

Olle Nerman

Göteborgs universitet

Chalmers, Matematiska vetenskaper, matematisk statistik

Linda Rörkvist

Göteborgs universitet

Anders Ståhlberg

Göteborgs universitet

Pierre Åman

Göteborgs universitet

PLoS ONE

1932-6203 (ISSN)

Vol. 7 e33208- e33208

Ämneskategorier

Cancer och onkologi

DOI

10.1371/journal.pone.0033208

PubMed

22496745