Nucleic Acids Research, Vol 24, Issue 24 4969-4977, Copyright © 1996 by Oxford University Press
P Ioannidis, M Havredaki, N Courtis and T Trangas
It has been demonstrated that the half-life of c-myc mRNA is modulated in
response to physiological agents. The elucidation of the decay process and
the identification of the critical steps in the in vivo c- myc mRNA
degradation pathway can be approached by following the fate of c-myc mRNA
under the influence of such factors. IFN-alpha was the factor used to
modulate c-myc mRNA half-life in HeLa 1C5 cells, a stable clone derived
from HeLa cells. This cell line carries multiple copies of the c-myc gene,
under the control of the dexamethasone inducible mouse mammary tumor
virus-long terminal repeat (MMTV-LTR). Exposure of HeLa 1C5 cells to
IFN-alpha resulted in a further 2-fold increase over the
dexamethasone-induced c-myc mRNA. However, the c-myc mRNA in IFN-alpha
treated cells was less stable than that in the control cells. RNase H
mapping of the 3' untranslated region of c-myc mRNA revealed, in addition
to the full length mRNA, three smaller fragments. These fragments were
proven to be truncated, non-adenylated c-myc mRNA species generated in
vivo. Exposure of HeLa 1C5 cells to Interferon-alpha before induction with
dexamethasone resulted in the enhanced presence of these intermediates.
RNase H analysis of c-myc mRNA after actinomycin D chase revealed that
deadenylation led to the formation of a relatively more stable
oligoadenylated c-myc mRNA population which did not appear to be precursor
to the truncated intermediates. The detection of truncated 3' end c-myc
mRNA adenylated fragments as well, implies that the c-myc mRNA degradation
process may follow an alternative pathway possibly involving
endonucleolytic cleavage.
ARTICLES
In vivo generation of 3' and 5' truncated species in the process of c- myc mRNA decay
Institute of Biology, NCSR-Demokritos, Athens, Greece.
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