The human MYOD1 transgene is suppressed by 5-bromodeoxyuridine in mouse myoblasts.
5-Bromodeoxyuridine (BrdU) immediately and clearly suppresses expression of the mouse Myod1 and human MYOD1 genes in myoblastic cells. Despite various studies, its molecular mechanism remains unknown. We failed to identify a BrdU-responsive element of the genes in experiments in which reporter constructs containing known regulatory sequences were transferred to mouse C2C12 myoblasts. Therefore, we transferred human chromosome 11 containing the MYOD1 gene to the cells by microcell-mediated chromosome transfer. In the resulting microcell hybrids, BrdU suppressed expression of the transgene, as determined by quantitative real-time RT-PCR analysis. We then transfected human PAC clones containing the MYOD1 gene to the cells. In the resulting transfectants, BrdU suppressed the transgene similarly. Deletion analysis suggested that a BrdU-responsive element or chromatin structure exists between 24 and 47 kb upstream of the gene. These results are the first demonstrating BrdU-responsiveness of a transgene for the known BrdU-responsive genes and facilitating determination of its precise responsible structure.
Ogino
Hideki
H
Kihara Institute for Biological Research and Graduate School of Integrated Science, Yokohama City University, Maioka-cho, Yokohama 244-0813, Japan.
Satou
Wataru
W
Fujii
Michihiko
M
Suzuki
Toshikazu
T
He
Yuan
Y
Michishita
Eriko
E
Ayusawa
Dai
D
D000818:Animals; D000963:Antimetabolites / Q000378:metabolism*; D001973:Bromodeoxyuridine / Q000378:metabolism*; D002478:Cells, Cultured; D002880:Chromosomes, Human, Pair 11; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D006824:Hybridization, Genetic; D051379:Mice; D017570:MyoD Protein / Q000235:genetics* / Q000378:metabolism; D032446:Myoblasts / Q000166:cytology / Q000502:physiology*; D012044:Regression Analysis; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D019076:Transgenes; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
D000963:Antimetabolites; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D015534:Trans-Activators; D014746:Vimentin; D001973:Bromodeoxyuridine