Microarray analysis detects novel Pax3 downstream target genes.
Pax3 is a transcription factor that is required for the development of embryonic neural tube, neural crest, and somatic derivatives. Our previous study (Mayanil, C. S. K., George, D., Mania-Farnell, B., Bremer, C. L., McLone, D. G., and Bremer, E. G. (2000) J. Biol. Chem. 275, 23259-23266) reveals that overexpression of Pax3 in a human medulloblastoma cell line, DAOY, resulted in an up-regulation in alpha-2,8-polysialyltransferase (STX) gene expression and an increase in polysialic acid on neural cell adhesion molecule. This finding suggests that STX might be a previously undescribed downstream target of Pax3. Because Pax3 is important in diverse cellular functions during development, we are interested in the identification of additional downstream targets of Pax3. We utilized oligonucleotide arrays and RNA isolated from stable Pax3 transfectants to identify potential target genes. A total of 270 genes were altered in the Pax3 transfectants as compared with the vector control and parental cell line. An independent analysis by cDNA expression array and real-time quantitative polymerase chain reaction of several genes confirmed the changes observed by the oligonucleotide microarray data. Of the genes that displayed significant changes in expression, several contain paired and homeodomain binding motifs of Pax3 in their promoter regions. Using promoter-luciferase reporter transfection assays and electromobility shift assays, we showed at least one previously undescribed downstream target, STX, to be a biological downstream target of Pax3. Thus we report several previously undescribed candidate genes to be potential downstream targets of Pax3.
Mayanil
C S
CS
Pediatric Brain Tumor Research Program, Department of Neurological Surgery, Northwestern University Medical School and Children's Memorial Institute of Education and Research, Chicago, Illinois 60614, USA.
George
D
D
Freilich
L
L
Miljan
E J
EJ
Mania-Farnell
B
B
McLone
D G
DG
Bremer
E G
EG
D000818:Animals; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D006801:Humans; D037181:Lectins, C-Type; D008527:Medulloblastoma; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D011401:Promoter Regions, Genetic; D012313:RNA / Q000378:metabolism; D015203:Reproducibility of Results; D012799:Sialyltransferases / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D053675:Versicans; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D011508:Chondroitin Sulfate Proteoglycans; D004268:DNA-Binding Proteins; D037181:Lectins, C-Type; D000072041:PAX3 Transcription Factor; C489409:PAX3 protein, human; D051761:Paired Box Transcription Factors; D014157:Transcription Factors; C508322:VCAN protein, human; C508323:Vcan protein, mouse; D053675:Versicans; C068686:Pax3 protein, mouse; D012313:RNA; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase