Identification of target genes regulated by PAX3 and PAX3-FKHR in embryogenesis and alveolar rhabdomyosarcoma.
PAX3 is a transcription factor important for neural, muscle, and facial development in vertebrates. To identify genes regulated by PAX3, we used a cyclic amplification and selection of targets (CASTing) strategy to isolate cis-regulatory elements bound by PAX3. CASTing libraries were constructed with mouse DNA fragments bound by mouse PAX3, and human genomic DNA fragments bound by human PAX3 and the fusion protein PAX3-FKHR. Approximately 1000 clones were sequenced from each of these three libraries. Numerous putative targets of PAX3 and PAX3-FKHR were identified and six genes, Itm2A, Fath, FLT1, TGFA, BVES, and EN2, were examined closely. The genomic DNA fragments near these genes contain PAX3 binding sites and confer PAX3-dependent regulation. The expression levels of these genes correlate with the PAX3 expression levels in mouse embryos or with PAX3-FKHR expression levels in rhabdomyosarcoma cell lines, and indicate they may be part of the PAX3 regulatory circuitry during embryogenesis and tumor formation.
Barber
Thomas D
TD
National Institute on Deafness and Other Communication Disorders, National Institutes of Health, 5 Research Court, Rockville, Maryland 20850, USA.
Barber
Melisa C
MC
Tomescu
Oana
O
Barr
Frederic G
FG
Ruben
Steve
S
Friedman
Thomas B
TB
D000818:Animals; D015815:Cell Adhesion Molecules / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D016326:Extracellular Matrix Proteins / Q000235:genetics; D000071161:Forkhead Box Protein O1; D051858:Forkhead Transcription Factors; D018507:Gene Expression Regulation, Developmental; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D018390:Gene Targeting; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics; D018995:Myosin Heavy Chains; D024501:Nonmuscle Myosin Type IIB; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D018232:Rhabdomyosarcoma, Alveolar / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D016211:Transforming Growth Factor alpha / Q000235:genetics; D014407:Tumor Cells, Cultured; D040281:Vascular Endothelial Growth Factor Receptor-1
C498789:Bves protein, mouse; D015815:Cell Adhesion Molecules; D004268:DNA-Binding Proteins; D016326:Extracellular Matrix Proteins; C085126:FOXO1 protein, human; D000071161:Forkhead Box Protein O1; D051858:Forkhead Transcription Factors; C467200:Foxo1 protein, mouse; C485527:ITM2A protein, human; C117398:Itm2a protein, mouse; D008565:Membrane Proteins; D009124:Muscle Proteins; D000072041:PAX3 Transcription Factor; C489409:PAX3 protein, human; D051761:Paired Box Transcription Factors; D014157:Transcription Factors; D016211:Transforming Growth Factor alpha; C068686:Pax3 protein, mouse; C501162:FLT1 protein, human; C501163:Flt1 protein, mouse; D040281:Vascular Endothelial Growth Factor Receptor-1; D024501:Nonmuscle Myosin Type IIB; C103815:nonmuscle myosin type IIB heavy chain; D018995:Myosin Heavy Chains