Figure 1. C. elegans strain VC2633 contains a mutation in
rpm-1(A) Representative images of PLM axon termination position in relation to ALM soma in L4 animals. ALM and PLM neurons are labelled with muIs32. Arrowheads indicate the axon end of PLM. Note that in
rpm-1(
ju1928)
degt-1(
ok3307) and
rpm-1(
ju1917) mutants, PLM axons terminate shortly past ALM soma, but do not display the hooking phenotype typically observed in
rpm-1(
ju44) and
rpm-1(
ju1918) mutants. (B) Quantification of the % of animals displaying PLM overextension phenotype, defined by at least one of the two PLM axons terminating past the ALM soma. n indicates the total number of animals quantified per genotype. Statistics was performed using Chi-squared test followed by Marascuilo procedure. #: significant, ns: not significant. (C) Genomic locus of
rpm-1 and
degt-1, and gene structure of
rpm-1 with the location of
ju1928 and
ju1917 adenine9267 (in CDS) changed to thymine.
ju1928 is linked to
degt-1(
ok3307) in strain VC2633.
ju1917 was generated by CRISPR/Cas9-mediated genome editing, and contains silent mutations near the a/t mutation to change restriction enzyme sites for the ease of genotyping.
ju1918 contains imprecise repair that causes insertion leading to premature stop, resulting in premature stop and truncated protein lacking C-terminal RING domain. (D) RPM-1 protein structure and domains: RCC1-like GEF domain (RLD), Pam, Highwire, RPM-1 family specific domains (PHR), RAE- 1 binding domain (RBD), FSN-1 binding domain (FBD1), B-Box Zn finger domains (B-Box) and RING-H2 ubiquitin ligase domain (RING). Below is a multi-species alignment (conserved residues are *) of the Q3089 region for C. elegans (C. e), H. sapiens (H. s.) and D. melanogaster (D. m.).