Distinct cytoplasmic and nuclear functions of the stress induced protein DDIT3/CHOP/GADD153
Journal article, 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.
Cell Movement
genetics
RNA
genetics
drug therapy
Antineoplastic Agents
drug therapy
Gene Expression Profiling
Tumor Markers
Liposarcoma
Cytoplasm
Messenger
genetics
Biological
Fibrosarcoma
drug effects
genetics
metabolism
Reverse Transcriptase Polymerase Chain Reaction
Cell Nucleus
Cell Adhesion
metabolism
genetics
Cultured
Recombinant Proteins
Humans
Genetic
metabolism
Fibroblasts
Tamoxifen
metabolism
metabolism
Oligonucleotide Array Sequence Analysis
metabolism
metabolism
drug effects
cytology
Hormonal
genetics
genetics
pharmacology
Green Fluorescent Proteins
Cells
Real-Time Polymerase Chain Reaction
Blotting
pharmacology
Cell Proliferation
genetics
drug effects
genetics
Cell Cycle
Western
Transcription Factor CHOP
Flow Cytometry
metabolism
Promoter Regions
Author
Alexandra Jauhiainen
Karolinska Institutet
Christer Thomsen
University of Gothenburg
Linda Strömbom
University of Gothenburg
Pernilla Grundevik
University of Gothenburg
Carola Andersson
University of Gothenburg
Anna Danielsson
University of Gothenburg
Mattias K Andersson
University of Gothenburg
Olle Nerman
University of Gothenburg
Chalmers, Mathematical Sciences, Mathematical Statistics
Linda Rörkvist
University of Gothenburg
Anders Ståhlberg
University of Gothenburg
Pierre Åman
University of Gothenburg
PLoS ONE
1932-6203 (ISSN)
Vol. 7 4 e33208- e33208Subject Categories (SSIF 2011)
Cancer and Oncology
DOI
10.1371/journal.pone.0033208
PubMed
22496745