Generation of a Moveable Poly(A)
            <sup>+</sup>
            Cassette

Cascone, Pamela J.; Schwartz, Lawrence M.

doi:10.2144/97231bm18

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“…The levels of polyubiquitin transcripts are barely detectable until the commitment to die, when transcript abundance increases over tenfold. The ubiquitin-fusion gene 80 (UBF80) represents the bulk of transcripts in the ISMs, in that their levels are not developmentally-regulated ed to p3′UB (to generate p3′UBAT) in order to examine the role of a poly A + tail on transcript properties (Cascone and Schwartz 1997).…”

Section: Figmentioning

confidence: 99%

“…To evaluate potential contributions by poly A + tails on message stability, an approximately 60-nucleotide-long poly A + sequence (Cascone and Schwartz 1997) was added to pLUB to generate pLUBAT. 35 S-labeled transcripts from pLUC (designated rLUC) and the four amended versions (rLA, rLF, rLUB, rLUBAT) were prepared and the t 1/2 of each associated mRNA determined in day 18 ISM extract (Fig.…”

Section: Developmental Changes In Mrna Stability In Ism Extractsmentioning

confidence: 99%

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Post-transcriptional regulation of gene expression during the programmed death of insect skeletal muscle

Cascone

Schwartz

2001

Development Genes and Evolution

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While considerable attention has focused on the role of specific proteins in mediating programmed cell death, few studies have examined the possible involvement of post-transcriptional regulation of mRNAs associated with this developmental process. We have examined developmental changes in transcript stability and translatability using protein extracts generated from the intersegmental muscles (ISM) of the moth Manduca sexta as a cell-free model system to examine three genes that are representative of the patterns of expression observed in condemned ISMs: repressed (actin), induced (polyubiquitin) and constitutively expressed (ubiquitin-fusion 80; ubf80). In addition, we have used luciferase mRNA as a generic reporter transcript to determine if there are sequence-specific controls of mRNA function related to programmed cell death. Among the three Manduca transcripts, polyubiquitin displayed the shortest half-life (t1/2) in all ISM extracts tested. The stability and translatability of all mRNAs were most affected in extracts from muscle cells from day 17 animals, just prior to the commitment of the muscles to die. Transfer of the 3' untranslated regions from the Manduca transcripts to luciferase mRNA did not appreciably change the stability or translatability of this test transcript. These data suggest that there may be global removal of cellular transcripts just prior to death to allow newly expressed mRNAs to rapidly accumulate to high levels. Such changes in message abundance, translatability and stability may facilitate the efficient activation of death (and perhaps other differentiation programs) in some developmental systems.

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Section: Figmentioning

confidence: 99%

Section: Developmental Changes In Mrna Stability In Ism Extractsmentioning

confidence: 99%