Specifically, by using mouse embryocarcinoma cells, we found that a dominant negative RARα mutant creates a concerted repression of both RARβ2 and its direct target CYP26A1, encoding the cytochrome P450 RA-specific hydrolase, which acts as a neuronal differentiation switch in these cells [8], [10].
Lastly, despite the power of yeast genetics, the interpretation of double or multiple mutant data is difficult as every mutant creates a pathological state and a function of a protein under these conditions may not necessarily reflect such a function in wild-type cells.
As such, antigenic mutants create a partially new niche and so do not necessarily lead to competitive exclusion.
A viable mutant created by insertion of a P-element (EP578) had been described as a suppressor of reaper-induced cell death [25], and a small deletion caused by excision of the P-element was reported to have generated a lethal null allele of the traf4 gene, traf4ex1 (formerly traf1ex1 [26]).
Electro-mobility shift assays (EMSAs) showed that the -151T mutant sequence created a high affinity binding site for a nuclear complex displaced by anti-Sp1 antibody and by competition with an Sp1 consensus oligonucleotides (Figure 4).
Each mutant created was confirmed by sequencing.
To determine if the lariat precursor remains the major pathway for circular RNA biogenesis in this mutant, we created an mrps16 mutant containing both deletion of the upstream 5′ ss and downstream branch point (to halt production of the lariat precursor), and, indeed, circular RNA production is strongly abated.
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To identify sites of anion interactions on α1 subunits, we used α1 mutants created from a long N-terminal isoform of CaV1.2 derived from rabbit cardiac cells.
The sorting out phenotype is consistent with the idea that the presence of mutant cells created an ectopic juxtaposition of cells with different identities.
