TY  - GEN
T1  - Three alleles in the pat-3 locus of Caenorhabditis elegans: mutations in the membrane-distal NPxY phosphotyrosine motif
AU  - Hanna, Jacob
AU  - Ramani, Shiva
AU  - Williams, Teja
AU  - Anaya, Ryan
AU  - Campion, Neil
AU  - Lopez, Evan
AU  - Williams, Raj
AU  - McIntire, Joe
AU  - Tran, Nicholas
AU  - Reyna, Victoria
AU  - Zeng, Jingmei
AU  - Miller, Shailyn
AU  - Pancar, Amar
AU  - Qiu, Zhongqiang
AU  - Lee, Myeongwoo
DO  - 10.17912/micropub.biology.000291
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000291/
AB  - Integrin is a heterodimeric cell surface receptor for extracellular matrix proteins. C. elegans has two α integrin and one β integrin subunit. The β integrin PAT-3 contains two NPxY phospho-tyrosine motifs in the cytoplasmic domain (Figure 1C). The NPxY motif is known for interacting with talin and kindlins and plays essential roles in the bidirectional signaling of integrins (Hynes 2002). To investigate the role of tyrosine phosphorylation in the NPxY motifs, we mutated the tyrosine to different amino acids to mimic the physiological modifications. In this study, the membrane-distal NPxY was studied using genome editing with the CRISPR-Cas9 ribonucleoprotein complex system (Dickinson and Goldstein 2016). The membrane distal 801NPVY804 was engineered to three different forms, such as NPVF804 (phenylalanine), NPVA804 (alanine), or NPVE804 (glutamate). The NPVF804 is to mimic the non-phosphorylatable tyrosine (Xu et al. 2010). NPVA804 is to abolish the tyrosine residue (Chen et al. 2006). NPVE804 is to mimic the phosphorylation (Qiu et al. 2019), with the expectation that three CRISPR engineered lines would display defective motility and abnormal cell migration. None of the lines, however, showed lethality or noticeable abnormal appearance, but they showed defective gonad migration (Figures 1B and 1D) and mild movement defects (Figure 1D). All alleles displayed a significant percentage of DTC migration defects (>30%) (Figure 1D). It should be noted that the DTC Mig was observed more frequently in the posterior gonad in kq8041 (NPVA804) and kq8042 (NPVE804), while the DTC Mig of kq8043 (NPVF804) was equally detected in both gonad arms. All alleles showed the decrease in motility; the kq8042 was severer than other alleles. We believe our results are useful for in vivo analysis of integrin functions and cell-matrix interactions.
PY  - 2020
JO  - microPublication Biology
ER  -