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Nucleic Acids Research 2004 32(16):4750-4757; doi:10.1093/nar/gkh808
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Published online 8 September 2004

Nucleic Acids Research, Vol. 32 No. 16 © Oxford University Press 2004; all rights reserved

The glioma-amplified sequence 41 gene (GAS41) is a direct Myb target gene

Daniel Braas, Holger Gundelach and Karl-Heinz Klempnauer*

Institut für Biochemie, Westfälische Wilhelms-Universität Münster, Wilhelm-Klemm Str.2, D-48149 Münster, Germany

* To whom correspondence should be addressed. Tel: +49 251 8333203; Fax: +49 251 8333206; Email: klempna{at}uni-muenster.de

Received as resubmission July 4, 2004; Accepted August 17, 2004

The retroviral oncogene v-myb encodes a transcription factor (v-Myb) which transforms myelomonocytic cells in vivo and in vitro. It is thought that v-Myb exerts its biological effects by deregulating the expression of specific target genes, most of which are still unknown. The chicken glioma-amplified sequence 41 gene (GAS41) is located immediately downstream of the lysozyme gene, a known Myb-regulated gene. The GAS41 promoter colocalizes with a CpG island which also functions as an origin of replication. Since the GAS41 promoter contains several potential Myb-binding sites (MBSs) we have investigated whether GAS41 is a v-Myb target gene. Our results show that the GAS41 gene is directly activated by a v-Myb/estrogen receptor fusion protein. Furthermore, our studies reveal that the GAS41 promoter is stimulated by v-Myb in co-transfection experiments and that the DNA-binding activity of v-Myb is crucial for transactivation of the promoter. Electrophoretic mobility-shift assays (EMSA) indicate that several Myb-binding sites, residing ~250 bp upstream of the transcriptional start site, are bound by Myb in vitro. Furthermore, chromatin immunoprecipitation assays demonstrate that v-Myb is bound to the GAS41 promoter in vivo. Taken together these findings identify the GAS41 gene as a novel v-Myb target gene. We have also analysed the GAS41 replication origin in myelomonocytic cells and have failed to observe significant differences in origin activity in cells expressing or not expressing v-Myb.


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