Spermine oxidase (SMO) is a flavoenzyme involved in polyamine homeostasis in animal cells. The mouse spermine oxidase gene (mSMO) codes for splice variants, including the previously reportedmajor active isoform, herein named alfa (a). In the present work, eight additional gene splicing variants were characterized. The heterologous expression and biochemical characterization ofthree recombinant isoforms (namely mSMOl, -c and -d) revealed that only the recombinant protein mSMOl displays biochemical characteristics similar to those ofm SMOa; the other two recombinant proteins contained no detectable SMO activity. In order to investigate in greater detail, the SMO enzyme activity associated with their subcellular localization,mSMOa and -l V5-tagged proteins were transiently and stably transfected in the murine neuroblastoma cell line, N18TG2. Very interestingly, the novel active mSMOl isoform was found to be present in both nuclear and cytoplasmic compartments, thus providing the first evidence of SMOactivity in the nucleus,while a cytoplasmic localization was confirmed for the mSMOa isoform. In addition, the relative transcription levels ofthe gene splicing variants were evaluated by RT-PCR analysis to verify a relationship with the SMO enzyme activity in various murine organs.
Cervelli, M., Bellini, A., Bianchi, M., Marcocci, L., Nocera, S., POLTICELLI F., F.R., et al. (2004). MOUSE SPERMINE OXIDASE GENE SPLICE VARIANTS. NUCLEAR SUBCELLULAR LOCALIZATION OF A NOVEL ACTIVE ISOFORM. EUROPEAN JOURNAL OF BIOCHEMISTRY, 271, 760-770 [10.1111/j.1432-1033.2004.03979.x].
MOUSE SPERMINE OXIDASE GENE SPLICE VARIANTS. NUCLEAR SUBCELLULAR LOCALIZATION OF A NOVEL ACTIVE ISOFORM
CERVELLI, MANUELA;
2004-01-01
Abstract
Spermine oxidase (SMO) is a flavoenzyme involved in polyamine homeostasis in animal cells. The mouse spermine oxidase gene (mSMO) codes for splice variants, including the previously reportedmajor active isoform, herein named alfa (a). In the present work, eight additional gene splicing variants were characterized. The heterologous expression and biochemical characterization ofthree recombinant isoforms (namely mSMOl, -c and -d) revealed that only the recombinant protein mSMOl displays biochemical characteristics similar to those ofm SMOa; the other two recombinant proteins contained no detectable SMO activity. In order to investigate in greater detail, the SMO enzyme activity associated with their subcellular localization,mSMOa and -l V5-tagged proteins were transiently and stably transfected in the murine neuroblastoma cell line, N18TG2. Very interestingly, the novel active mSMOl isoform was found to be present in both nuclear and cytoplasmic compartments, thus providing the first evidence of SMOactivity in the nucleus,while a cytoplasmic localization was confirmed for the mSMOa isoform. In addition, the relative transcription levels ofthe gene splicing variants were evaluated by RT-PCR analysis to verify a relationship with the SMO enzyme activity in various murine organs.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
