FIGURE

FIGURE 3

ID

ZDB-FIG-211013-3

Publication

Pakdaman et al., 2021 - Chip Protein U-Box Domain Truncation Affects Purkinje Neuron Morphology and Leads to Behavioral Changes in Zebrafish

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FIGURE 3

Figure 3. Zebrafish U-box^{− /−} expressing truncated Chip with impaired ubiquitin ligase activity were generated by CRISPR/Cas9. Zebrafish stub1 gene was targeted for mutagenesis at the last exon (A), leading to an introduction of 7 bp-deletion mutation in this exon (c.761-769del) which was detected by MluI restriction digestion (B) and DNA sequencing (C). The 761-769del mutation is predicted to result in a frameshift and early stop codon and truncation of C-terminal U-box domain (D). Recombinant Chip proteins were produced from the cDNA of U-box^{− /−} and wild-type zebrafish, and further examined for in vitro ubiquitin ligase activity on Chip (E, lane 1–3) and Hsc70 substrates (E, lane 4–6). Compared to the wild-type human CHIP (E, lane 1 and 4) and wild-type zebrafish Chip (E, lane 2 and 5) controls, U-box truncated zebrafish Chips showed impaired self-ubiquitination (E, lane 3) and Hsc70 ubiquitination activities (E, lane 6).

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Front. Mol. Neurosci.