Activation of human c-raf-1 by replacing the N-terminal region with different sequences

Abstract

Two transformants of NIH 3T3 cells, obtained by the transfection of human colon cancer and normal colon DNAs, contained activated c- raf -1. In both the activated c- raf -1, the 5′ half of the c- raf -1 sequence was replaced by sequences other than c- raf -1 as a result of recombinations which occurred at the intron between exons 7 and 8. It was suggested, however, that these recombinations, which conferred the transforming activity on the c- raf -1, occurred during the transfection. In one case analyzed, characteristic sequences were found near the breakpoint and these may be involved in the recombination. It was found, upon analysing the structure of the cDNA derived from one of the activated c- raf -1, that fused mRNA had been transcribed from the recombined gene comprising the non- raf gene and c- raf -1. The mRNA possibly encodes a fused protein. One cDNA clone was derived from alternatively spliced mRNA, although its physiological role is unclear. On comparing the structure of the two human activated c- raf -1 and the rat activated c- raf which we have reported previously (5), it was revealed that, in these three cases, the sequences joined to the truncated c- raf (−1) ¹ were different. It was suggested from data which we and others have previously reported that various sequences could be capable of activating c- raf (−1) by replacing its 5′ half.