Expression of the mRNA (N6-adenosine)-methyltransferase S-adenosyl-L-methionine binding subunit mRNA in cultured...

Ribonuclease protection assays (RPA) were used to detect and quantitate the amount of messenger RNA (mRNA) coding for the S-adenosyl-L-methionine binding subunit (MT-A70) of the mRNA (N6-adenosine)-methyltransferase from different types of cultured cells. HeLa cells cultured in suspension were analyzed at regular intervals along a normal growth curve. It was discovered that MT-A70 mRNA was transcribed constitutively across the time-course, irrespective of the rate of cellular proliferation. Further, 11 different cell lines representing non-tumorigenic, tumorigenic, and virally-transformed tumorigenic types from Homo sapiens, Mus musculus, and Rattus norvegicus were examined for MT-A70 mRNA expression. It was found that all the cell lines expressed a long and short splice-variant form of the gene. In general, the cell lines expressed a similar total amount of the MT-A70 mRNA while statistically significant differences existed between the quantity of the long and short forms among cell types. Tumorigenic cell lines synthesized as much as a 9-fold greater amount of long form versus short form MT-A70 mRNA. Comparatively, non-tumorigenic cell lines generally expressed only a 1.5-fold greater amount of long form versus short form MT-A70 mRNA.