Nucleic Acids Research, 1988, Vol. 16, No. 16 8029-8044
© 1988
Articles |
Methylated DNA-binding protein is present in various mammalian cell types
Department of Biochemistiy, Tulane Medical School New Orleans, LA 70112, USA 1Southern Regional Research Center, US Department of Agriculture New Orleans, LA 70122, USA
Received February 29, 1988. Revised July 12, 1988. Accepted July 12, 1988.
A DNA-binding protein from human placenta, methylated DNA-binding protein (MDBP), binds to certain DNA sequences only when they contain 5-methylcytosine (m5C) residues at specific positions. We found a very similar DNA-binding activity in nuclear extracts of rat tissues, calf thymus, human embryonal carcinoma cells, HeLa cells, and mouse LTK cells. Like human placental MDBP, the analogous DNA-binding proteins from the above mammalian cell lines formed a number of different low-electrophoretic-mobility complexes with a 14-bp MDBP-specific oligonucleotide duplex. All of these complexes exhibited the same DNA methylation specificity and DNA sequence specificity. From the extracts of rat and calf tissues, oligo-nucleotide protein complexes formed that also had the same specificity as human placental MDBP although they had a higher electrophoretic mobility probably due to digestion by proteases in the nuclear extract8. Although MDBP activity was found in various mRn1mi cell types, it was not detected in extracts of cultured mosquito cells and so may be associated only with cells with vertebrate-type DNA methylation.