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Messenger RNAs were further isolated by binding to a polyA column.
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Npl3 was observed to compete strongly for binding to the polyA signal (Figure 4A).
Rna15 was not phosphorylated by Cka1, nor was binding to the polyA signal affected in reactions with Cka1 lacking Npl3 (data not shown).
In addition, Casein Kinase 2 was found to be required for the phosphorylation of Npl3 and affect its ability to compete against Rna15 (Cleavage Factor I) for binding to polyA signals.
First, the N‐terminal zinc finger of Pan3 conveys sequence specificity by binding to polyA RNA preferentially over other polyribonucleotides (Supplementary Fig S4).
As a reverse primer we used 3' GeneRacer oligo attached to a polyA tail during first-strand cDNA synthesis.
These include observations showing that ataxin-2 is a component of the polysome complex and that it binds to polyA binding protein 1 (PABP-1) in translation initiation [8].
One of miR functions is promotion of mRNA decay by directing polyA tail removal via incomplete complementary binding to mRNA [30].
A model where Msi proteins repress translation by outcompeting eIF4G for PolyA-binding protein (PABP) was proposed (Kawahara et al., 2008), but the conditions under which binding to mRNA results in translational repression are unclear, since only a subset of mRNAs are detectably regulated.
This defect is due to mutations in RRM2, which result in reduced the binding specificity of Npl3 [42], [42], and reduce the ability of the protein to compete effectively for binding to polyA/termination sequences [12].
The alternative, less abundant PDCD2 mRNA is formed due to a polyadenylation (polyA) signal in the third intron.
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