To confirm that capt was the affected gene, we sequenced the capt locus from the punctate mutant and found a nonsense mutation that changed amino acid lysine 304 to a premature stop codon.
To determine the molecular lesion responsible for this phenotype, we used deficiency mapping and identified two molecularly defined deficiencies, Df 2L Exel7006 and Df(2l Exel6004, that failed to rescue the lethality of the punctate mutant.
These actin::GFP puncta, which are absent in wild type larvae, occur at an average of 38 puncta (standard deviation = 10) in each sensory neuron dorsal cluster of the punctate mutant.
We found that the GFP signal was enhanced in the punctate regions in unc-13 mutants, as in unc-2 mutants.
These data show that intracellular polymers contribute to the punctate accumulations of mutant neuroserpin in cell models of FENIB [(27) and the present work], and together with our ELISA results shown above demonstrate that our monoclonal antibodies are a valuable tool for the detection and quantitation of total and polymerized neuroserpin.
(C ) PTRN-1a ΔCKK)::tdTomato PTRN-1a ΔCKK tdTomato-1(tm5597) mutant expressedpunctate localinathen in the ptrn-1 tm5597the ptrn-1 tm5597
Moreover, when PIASy was coexpressed, the K171R mutant dramatically localized to the nucleus but lost the punctate foci which overlapped with PIASy in 786-O cells (Figure 3A and B, left panels).
This pattern of colocalization supports the idea that the punctate Notch staining found in Hpo mutant follicle cells reflects the accumulation of Notch in endocytic vesicles.
Although, the Ku70-TRF1 fluorescence was decreased in the presence of the TRF2ΔBΔM mutant, the signal remained punctate.
In unc-2 mutants, we found that the GFP signal is significantly enriched within the punctate regions and the GFP RAB-3 puncta are larGFP RAB-3righter compunctato wild type.
