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Vol. 10, Issue 3, 515-524, March 1999
Departments of Immunology and Molecular and Medical Genetics,
University of Toronto, Toronto, Ontario, Canada M5S 1A8
In many organisms nonsense mutations decrease the level of mRNA. In
the case of mammalian cells, it is still controversial whether
translation is required for this nonsense-mediated RNA decrease (NMD).
Although previous analyzes have shown that conditions that impede
translation termination at nonsense codons also prevent NMD, the
residual level of termination was unknown in these experiments. Moreover, the conditions used to impede termination might also have
interfered with NMD in other ways. Because of these uncertainties, we
have tested the effects of limiting translation of a nonsense codon in
a different way, using two mutations in the immunoglobulin µ heavy
chain gene. For this purpose we exploited an exceptional nonsense
mutation at codon 3, which efficiently terminates translation but
nonetheless maintains a high level of µ mRNA. We have shown 1) that
translation of Ter462 in the double mutant occurs at only ~4% the
normal frequency, and 2) that Ter462 in cis with Ter3 can induce NMD. That is, translation of Ter462 at this low (4%) frequency is sufficient to induce NMD.
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