Discovery of Independent Suppressors in C. elegans During CMTR1 Knockdown: Insights from a Generic Screen

Abstract

In this study, we used C. elegans as a model organism to investigate the regulation and function of CMTR1 through a direct mutagenesis screen using ethyl methanesulfonate (EMS). Our goal was to identify suppressors of CMTR1 knockdown that could restore fertility, employing an auxin-inducible degradation (AID) system to target CMTR1. However, we anticipated that EMS mutagenesis would reveal faults in the AID system, particularly through TIR1 loss-of-function mutations, which inadvertently prevented CMTR1 degradation and allowed the variants to maintain fertility. Our results confirmed our hypothesis, showing that TIR1 loss-of-function mutations prevent the degradation of CMTR1, thereby allowing CMTR1 to persist and the variants to maintain fertility. Analysing the frequency and nature of these mutations will be essential for estimating the size of the genuine suppressor targets identified through EMS screening. This approach will help us gauge the extent and scope of the suppressors that can compensate for CMTR1 knockdown. In addition to these control-related findings, we successfully identified Suppressors of CMTR1 Knockdown (SOCK) that restored fertility through mutations in other proteins or regulatory sequences. These findings suggest a complex regulatory network involving CMTR1, highlighting its crucial function beyond immune recognition and opening new avenues for understanding the broader biological roles of 2'-O-ribose methylation. In conclusion, this research can provide a clearer understanding of the importance of cap 1 and 2'-O-ribose methylation in cellular activities, highlighting their crucial roles in preserving fertility