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To determine if the TERT TEN domain mutants localize to telomeres, the authors overexpress the mutants and examined localization to telomeres or Cajal bodies.
Strikingly, although not surprisingly, these mutants localize to the bisdioxopiperazine binding site in the N-terminal domain of TOP2 [ 26].
Since TEN-domain mutants localize to Cajal bodies and not some other structure, there is no reason to believe that there is an assembly or maturation defect caused by overexpression.
For example, identification of the interfaces of the aberrant oligomers helps explain why Cu2Zn2SOD1S S is so resistant to oligomerization, and potentially, why many of the ALS mutants localize to the dimer or the metal-binding region.
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Only the Y646F mutant co-localized with nuclear pore complex proteins, including Nup53 and Nup98, whereas the other mutants localized to the nuclear envelope rim.
In addition, the shs1-ps mutants localized to the bud neck (Figure S1).
In addition, Cdc14A phosphorylation mutants localized to centrosomes as did wt-Cdc14A (Suppl. Fig. S2A and data not shown).
With the exception of the S418A mutant, all other mutants localized to centrosomes and to spindle microtubules, and supported the targeting of γ-tubulin to these structures, similar to the wild type protein.
This information was required before we could compile lists of cloned emb mutants that were logical candidates for allelism tests with mapped emb mutants localized to similar regions of the genome.
All the missense mutants localized to similar subcellular regions as the corresponding WT proteins (Fig. 5B).
All mutants localized to the plasma membrane, with minimal evidence of intracellular staining (Fig. 3A).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com