Hi All,
Could anyone give me some suggestion as to what are the best assays for determine the oncogenic domain on a oncoprotein?
Currently I am working on a protein that known to be upregulated in many different cancers and should be a oncogene. However, since it is a multidomain protein, we are not sure which domain contribute most in its oncogenic property.
I know that NIH3T3 cell transformation assay could be an option, but are there other assays that are also quite frequently used for this purpose? (I prefer cell biology assays not animal assay because the animal assay needs lot more $$$)
Thank you,
Lyra
If protein of interest contains domains that are known and well-characterized, it should be possible to predict its activity such as transcription factor, adaptor, or receptor etc. This information is sufficient to design experiments to test which domain in the protein is required for its oncogenic activity in the classical NIH 3T3 cell transformation assay.
Hello Lyra,
besides the good suggestion from Divaker, you could also try to do some data mining analysis by checking if your gene is also activated through the presence of, say, point mutations (for instance you could have a look at the cosmic website: http://cancer.sanger.ac.uk/cosmic). Quite often, activating point mutations cluster in very narrow regions of an oncogene, usually highlighting the presence of critical domains.
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