Polyamine-mediated regulation of mouse ornithine decarboxylase is posttranslational.
The activity of ornithine decarboxylase (ODC) is negatively regulated by intracellular polyamines, which thereby mediate a form of feedback inhibition of the initial enzyme in the pathway of their synthesis. This phenomenon has been believed to result, at least in part, from translational regulation. To investigate this further, we performed four series of experiments. First, we found that a chimeric protein encoded by an mRNA containing the ODC 5' leader sequence did not exhibit polyamine-dependent regulation. Second, we showed that transcripts containing the protein-coding sequence of ODC, but no other ODC-derived sequence information, exhibited regulation. Third, we found that the association of ODC mRNA with ribosomes was not altered when intracellular polyamine levels were modulated under conditions previously deemed to cause translational regulation. Last, we carried out experiments to measure the incorporation of [35S]methionine into ODC in polyamine-starved and polyamine-replete cells. Differential incorporation diminished progressively as pulse-label times were shortened; at the shortest labeling time used (4 min), the difference in favor of ODC in polyamine-starved cells was less than twofold. These findings suggest that it is necessary to reevaluate the question of whether polyamines cause alterations of translation of ODC mRNA.
van Daalen Wetters
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Department of Microbiology and Immunology, University of California, San Francisco 94143.
Macrae
M
M
Brabant
M
M
Sittler
A
A
Coffino
P
P
D000199:Actins / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D005796:Genes; D051379:Mice; D008815:Mice, Inbred Strains; D009955:Ornithine Decarboxylase / Q000096:biosynthesis / Q000235:genetics*; D011132:Polyribosomes / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011700:Putrescine / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D000199:Actins; D004274:DNA, Recombinant; D012333:RNA, Messenger; D001616:beta-Galactosidase; D009955:Ornithine Decarboxylase; D011700:Putrescine