Activation of the mas oncogene involves coupling to human alphoid sequences.
We have shown previously that mouse NIH3T3 cells transfected with DNA from a human ovarian carcinoma were rendered tumourigenic by an activated mas oncogene in four independent transfection experiments. In all cases the 5'-noncoding region was rearranged in comparison to the original ovarian tumour DNA. We now report that in all four transfectants the newly acquired sequences consist of human centromeric alpha satellite repeat DNA. In at least three transfectants the alphoid DNA originates from the centromere of chromosome three. Analysis of the sequences of the recombination site in one transfectant revealed that a homologous sequence of five base pairs (CAGCA) is present in both parental strands, and might thus have contributed to the recombinational event. To establish a conclusive role for alphoid DNA in the activation of mas, we performed a co-transfection experiment in NIH3T3 cells with cloned alphoid DNA and the mas coding sequence. We show that the transfectants expressing a transformed phenotype contain amplified mas linked to alphoid DNA. NIH3T3 cells transfected with plasmids that contained alphoid sequences cloned directly upstream of the mas coding sequence, and injected into nude mice, gave rise to tumours with amplified mas sequences (7/7). In six of these tumours the alphoid sequences were amplified as well. Our data suggest a novel mechanism of oncogene activation: recombination with normal alphoid repeat DNA resulting in amplification of the oncogene.
Department of Clinical Oncology, University Hospital, Leiden, The Netherlands.
L J
LJ
van 't Veer
M J
MJ
van der Feltz
C A
CA
van den Berg-Bakker
N C
NC
Cheng
R P
RP
Hermens
D A
DA
van Oorschot
T
T
Kievits
P I
PI
Schrier
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002503:Centromere; D002893:Chromosomes, Human, Pair 3; D004273:DNA, Neoplasm / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics* / Q000502:physiology; D008040:Genetic Linkage; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics*; D009368:Neoplasm Transplantation; D009857:Oncogenes / Q000235:genetics* / Q000502:physiology; D010051:Ovarian Neoplasms / Q000235:genetics*; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D043562:Receptors, G-Protein-Coupled; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014162:Transfection
D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D043562:Receptors, G-Protein-Coupled