Amplification of nuclear aldolase A in mouse cell mutants resistant to Hoechst 33342.
5-Bromodeoxyuridine (BrdU) induces a phenomenon similar to cellular senescence in mammalian cells. AT-binding ligands such as Hoechst 33258 synergistically potentiate the effect of BrdU. We isolated mouse FM3A cell mutants resistant to Hoechst 33342 and characterized two highly resistant mutants. Two-dimensional gel electrophoresis followed by peptides sequence tags revealed that nuclear aldolase A was markedly increased in both mutants. Western blot analysis confirmed that nuclear aldolase A was increased leaving cytosolic aldolase A unaltered. Its mRNA levels were also increased in the mutants. Enforced expression of aldolase A conferred resistance to Hoechst 33342 on wild-type cells. Taken together, nuclear aldolase A was shown to somehow protect the cytotoxic effect of Hoechst 33342.
Kihara Institute for Biological Research and Graduate School of Integrated Science, Yokohama City University, Maioka-cho 641-12, Yokohama 244-0813, Japan.
Wataru
W
Satou
Hiroshi
H
Tanimoto
Ryo
R
Ukekawa
Michihiko
M
Fujii
Dai
D
Ayusawa
D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology*; D006690:Bisbenzimidazole / Q000494:pharmacology; D015153:Blotting, Western; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000201:enzymology*; D003600:Cytosol / Q000378:metabolism; D004351:Drug Resistance; D015180:Electrophoresis, Gel, Two-Dimensional; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology; D051379:Mice; D009154:Mutation; D009687:Nuclear Proteins / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D014162:Transfection
D001562:Benzimidazoles; D007211:Indoles; D009687:Nuclear Proteins; D012333:RNA, Messenger; C007293:DAPI; D005634:Fructose-Bisphosphate Aldolase; D001973:Bromodeoxyuridine; D006690:Bisbenzimidazole; C017807:bisbenzimide ethoxide trihydrochloride