Upstream ORF affects MYCN translation depending on exon 1b alternative splicing

<p>Abstract</p> <p>Background</p> <p>The <it>MYCN </it>gene is transcribed into two major mRNAs: one full-length (<it>MYCN) </it>and one exon 1b-spliced (<it>MYCN</it>^{Δ1<it>b</it>}) mRNA. But nothing is known about their respective ability to translate the MYCN protein.</p> <p>Methods</p> <p>Plasmids were prepared to enable translation from the upstream (uORF) and major ORF of the two <it>MYCN </it>transcripts. Translation was studied after transfection in neuroblastoma SH-EP cell line. Impact of the upstream AUG on translation was evaluated after directed mutagenesis. Functional study with the two <it>MYCN </it>mRNAs was conducted by a cell viability assay. Existence of a new protein encoded by the <it>MYCN</it>^{Δ1<it>b </it>}uORF was explored by designing a rabbit polyclonal antibody against a specific epitope of this protein.</p> <p>Results</p> <p>Both are translated, but higher levels of protein were seen with <it>MYCN</it>^{Δ1<it>b </it>}mRNA. An upstream ORF was shown to have positive cis-regulatory activity on translation from <it>MYCN </it>but not from <it>MYCN</it>^{Δ1<it>b </it>}mRNA. In transfected SH-EP neuroblastoma cells, high MYCN dosage obtained with <it>MYCN</it>^{Δ1<it>b </it>}mRNA translation induces an antiapoptotic effect after serum deprivation that was not observed with low MYCN expression obtained with <it>MYCN </it>mRNA. Here, we showed that MYCNOT: <it>MYCN </it>Overlap Transcript, a new protein of unknown function is translated from the upstream AUG of <it>MYCN</it>^{Δ1<it>b </it>}mRNA.</p> <p>Conclusions</p> <p>Existence of upstream ORF in <it>MYCN </it>transcripts leads to a new level of MYCN regulation. The resulting MYCN dosage has a weak but significant anti-apoptotic activity after intrinsic apoptosis induction.</p

Complex kame belt morphology, stratigraphy and architecture

The development of glacier karst at the margins of melting ice sheets produces complex glaciofluvial sediment-landform assemblages that provide information on ice sheet downwasting processes. We present the first combined geomorphological, sedimentological and geophysical investigation of the Brampton Kame Belt, an important glaciofluvial depositional zone at the centre of the last British-Irish Ice Sheet. Ground-penetrating radar (GPR) data allow the broad scale internal architecture of ridges (eskers) and flat-topped hills (ice-walled lake plains) to be determined at four sites. In combination with sediment exposures, these provide information on lateral and vertical variations in accretion styles, depositional boundaries, and grain size changes. Building on existing work on the subject, we propose a refined model for the formation of ice-walled lake plains resulting from the evolution and collapse of major drainage axes into lakes as stable glacier karst develops during deglaciation. The internal structure of esker ridges demonstrates variations in sedimentation that can be linked to differences in ridge morphologies across the kame belt. This includes low energy flow conditions and multiple accretion phases identified within large S-N oriented esker ridges; and fluctuating water pressures, hyperconcentrated flows, and significant deformation within a fragmented SW-NE oriented esker ridge. In combination with updated geomorphological mapping, this work allows us to identify two main styles of drainage within the kame belt: (1) major drainage axes aligned broadly S-N that extend through the entire kame belt and collapsed into a chain of icewalled lakes; and (2) a series of smaller, fragmented SW-NE aligned esker ridges that represent ice-marginal drainage as the ice sheet receded south-eastwards up the Vale of Eden. Our study demonstrates the importance of integrated geomorphological, sedimentological and geophysical investigations in order to understand complex and polyphase glaciofluvial sediment-landform assemblages