title: Dihydroorotate dehydrogenase mRNA and protein expression analysis in normal and drug-resistant cells.
abstract: To follow the expression of the fourth enzyme of pyrimidine de novo synthesis dihydroorotate dehydrogenase (DHODH) in cells and tissues, we studied the DHODH mRNA expression by means of RT-PCR in rat tissues. Rabbit polyclonal anti-DHODH immunoglobulins were applied for immunochemical quantification of the enzyme protein by Western blotting. In mouse B-lymphocytes, which were adapted to tolerate up to a 50-fold concentration of the DHODH inhibitor leflunomide, a 20 fold protein overexpression was measured. Southern blotting indicated DHODH gene amplification.
authors.affiliation: Institute for Physiological Chemistry, Philipps-University Marburg, Marburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Löffler
authors.forename: A
authors.initials: A
authors.lastname: Klein
authors.forename: M
authors.initials: M
authors.lastname: Hayek-Ouassini
authors.forename: W
authors.initials: W
authors.lastname: Knecht
authors.forename: L
authors.initials: L
authors.lastname: Konrad
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D004247:DNA / Q000737:chemistry; D000090722:Dihydroorotate Dehydrogenase; D004351:Drug Resistance; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D007150:Immunohistochemistry; D020128:Inhibitory Concentration 50; D007555:Isoxazoles / Q000494:pharmacology; D000077339:Leflunomide; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D008566:Membranes / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000378:metabolism*; D011743:Pyrimidines / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis; D013997:Time Factors
chemical_list: D000090722:Dihydroorotate Dehydrogenase; D007136:Immunoglobulins; D007555:Isoxazoles; D011743:Pyrimidines; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004247:DNA; D044925:Oxidoreductases Acting on CH-CH Group Donors; D000077339:Leflunomide