Source Publication
G3: Genes, Genomes, Genetics
Document Type
Article
Publication Date
2-2018
DOI
10.1534/g3.117.300298
Abstract
Separase is a highly conserved protease required for chromosome segregation. Although observations that separase also regulates membrane trafficking events have been made, it is still not clear how separase achieves this function. Here, we present an extensive ENU mutagenesis suppressor screen aimed at identifying suppressors of sep-1(e2406), a temperature-sensitive maternal effect embryonic lethal separase mutant that primarily attenuates membrane trafficking rather than chromosome segregation. We screened nearly a million haploid genomes and isolated 68 suppressed lines. We identified 14 independent intragenic sep-1(e2406) suppressed lines. These intragenic alleles map to seven SEP-1 residues within the N-terminus, compensating for the original mutation within the poorly conserved N-terminal domain. Interestingly, 47 of the suppressed lines have novel mutations throughout the entire coding region of the pph-5 phosphatase, indicating that this is an important regulator of separase. We also found that a mutation near the MEEVD motif of HSP-90, which binds and activates PPH-5, also rescues sep-1(e2406) mutants. Finally, we identified six potentially novel suppressor lines that fall into five complementation groups. These new alleles provide the opportunity to more exhaustively investigate the regulation and function of separase.
Recommended Citation
Melesse, Michael, Dillon E. Sloan, Joseph T. Benthal, Quincey Caylor, Krishen Gosine, Xiaofei Bai, and Joshua N. Bembenek. “Genetic Identification of Separase Regulators in Caenorhabditis elegans.” G3: Genes, Genomes, Genetics 8, no. 2 (2018). https://doi.org/10.1534/g3.117.300298.
Submission Type
Publisher's Version
Comments
This article was published openly thanks to the University of Tennessee Open Publishing Support Fund.
Licensed under a Creative Commons Attribution 4.0 International license.