title: Systematic analyses of rpm-1 suppressors reveal roles for ESS-2 in mRNA splicing in Caenorhabditis elegans.
abstract: The PHR (Pam/Highwire/RPM-1) family of ubiquitin E3 ligases plays conserved roles in axon patterning and synaptic development. Genetic modifier analysis has greatly aided the discovery of the signal transduction cascades regulated by these proteins. In Caenorhabditis elegans, loss of function in rpm-1 causes axon overgrowth and aberrant presynaptic morphology, yet the mutant animals exhibit little behavioral deficits. Strikingly, rpm-1 mutations strongly synergize with loss of function in the presynaptic active zone assembly factors, syd-1 and syd-2, resulting in severe locomotor deficits. Here, we provide ultrastructural evidence that double mutants, between rpm-1 and syd-1 or syd-2, dramatically impair synapse formation. Taking advantage of the synthetic locomotor defects to select for genetic suppressors, previous studies have identified the DLK-1 MAP kinase cascade negatively regulated by RPM-1. We now report a comprehensive analysis of a large number of suppressor mutations of this screen. Our results highlight the functional specificity of the DLK-1 cascade in synaptogenesis. We also identified two previously uncharacterized genes. One encodes a novel protein, SUPR-1, that acts cell autonomously to antagonize RPM-1. The other affects a conserved protein ESS-2, the homolog of human ES2 or DGCR14. Loss of function in ess-2 suppresses rpm-1 only in the presence of a dlk-1 splice acceptor mutation. We show that ESS-2 acts to promote accurate mRNA splicing when the splice site is compromised. The human DGCR14/ES2 resides in a deleted chromosomal region implicated in DiGeorge syndrome, and its mutation has shown high probability as a risk factor for schizophrenia. Our findings provide the first functional evidence that this family of proteins regulate mRNA splicing in a context-specific manner.
authors.affiliation: Neurobiology Section, Division of Biological Sciences, University of California, San Diego, La Jolla, California 92093 Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California 92093.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Noma
authors.affiliation: Neurobiology Section, Division of Biological Sciences, University of California, San Diego, La Jolla, California 92093 Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California 92093.
authors.forename: Alexandr
authors.initials: A
authors.lastname: Goncharov
authors.affiliation: Neurobiology Section, Division of Biological Sciences, University of California, San Diego, La Jolla, California 92093 Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California 92093 yijin@ucsd.edu.
authors.forename: Yishi
authors.initials: Y
authors.lastname: Jin
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D017238:Genes, Helminth; D016153:Genes, Suppressor; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics* / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D017729:Presynaptic Terminals / Q000378:metabolism / Q000648:ultrastructure; D022821:RNA Splice Sites / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism
chemical_list: D029742:Caenorhabditis elegans Proteins; D020662:Guanine Nucleotide Exchange Factors; D022821:RNA Splice Sites; D012333:RNA, Messenger; C409634:RPM-1 protein, C elegans
keywords: DLK-1 MAPKKK cascade; ESS-2/DGCR14; mRNA splicing; rpm-1; synapse development