Characterization of the c-MYC-regulated transcriptome by SAGE: identification and analysis of c-MYC target genes.
To identify target genes of the oncogenic transcription factor c-MYC, serial analysis of gene expression (SAGE) was performed after adenoviral expression of c-MYC in primary human umbilical vein endothelial cells: 216 different SAGE tags, corresponding to unique mRNAs, were induced, whereas 260 tags were repressed after c-MYC expression (P < 0.05). The induction of 53 genes was confirmed by using microarray analysis and quantitative real-time PCR: among these genes was MetAP2/p67, which encodes an activator of translational initiation and represents a validated target for inhibition of neovascularization. Furthermore, c-MYC induced the cell cycle regulatory genes CDC2-L1, Cyclin E binding protein 1, and Cyclin B1. The DNA repair genes BRCA1, MSH2, and APEX were induced by c-MYC, suggesting that c-MYC couples DNA replication to processes preserving the integrity of the genome. MNT, a MAX-binding antagonist of c-MYC function, was up-regulated, implying a negative feedback loop. In vivo promoter occupancy by c-MYC was detected by chromatin immunoprecipitation for CDK4, Prohibitin, MNT, Cyclin B1, and Cyclin E binding protein 1, showing that these genes are direct c-MYC targets. The c-MYC-regulated genes/tags identified here will help to define the set of bona fide c-MYC targets and may have potential therapeutic value for inhibition of cancer cell proliferation, tumor-vascularization, and restenosis.
Menssen
Antje
A
Max-Planck-Institute of Biochemistry, Molecular Oncology, Independent Junior Research Group, Am Klopferspitz 18A, D-82152 Martinsried/Munich, Germany.
Hermeking
Heiko
H
D000256:Adenoviridae / Q000235:genetics; D000483:Alleles; D001402:B-Lymphocytes / Q000378:metabolism; D015152:Blotting, Northern; D002453:Cell Cycle; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D015723:Gene Library; D016259:Genes, myc / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008957:Models, Genetic; D018919:Neovascularization, Physiologic; D020411:Oligonucleotide Array Sequence Analysis; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014471:Umbilical Veins / Q000166:cytology
D002843:Chromatin