Allele-specific mitochondrial stress induced by Multiple Mitochondrial Dysfunctions Syndrome 1 pathogenic mutations modeled in Caenorhabditis elegans

PLoS Genet. 2021 Aug 27;17(8):e1009771. doi: 10.1371/journal.pgen.1009771. eCollection 2021 Aug.

Abstract

Multiple Mitochondrial Dysfunctions Syndrome 1 (MMDS1) is a rare, autosomal recessive disorder caused by mutations in the NFU1 gene. NFU1 is responsible for delivery of iron-sulfur clusters (ISCs) to recipient proteins which require these metallic cofactors for their function. Pathogenic variants of NFU1 lead to dysfunction of its target proteins within mitochondria. To date, 20 NFU1 variants have been reported and the unique contributions of each variant to MMDS1 pathogenesis is unknown. Given that over half of MMDS1 individuals are compound heterozygous for different NFU1 variants, it is valuable to investigate individual variants in an isogenic background. In order to understand the shared and unique phenotypes of NFU1 variants, we used CRISPR/Cas9 gene editing to recreate exact patient variants of NFU1 in the orthologous gene, nfu-1 (formerly lpd-8), in C. elegans. Five mutant C. elegans alleles focused on the presumptive iron-sulfur cluster interaction domain were generated and analyzed for mitochondrial phenotypes including respiratory dysfunction and oxidative stress. Phenotypes were variable between the mutant nfu-1 alleles and generally presented as an allelic series indicating that not all variants have lost complete function. Furthermore, reactive iron within mitochondria was evident in some, but not all, nfu-1 mutants indicating that iron dyshomeostasis may contribute to disease pathogenesis in some MMDS1 individuals.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alleles
  • Animals
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans Proteins / genetics
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Disease Models, Animal
  • Iron / metabolism
  • Iron-Sulfur Proteins / genetics*
  • Iron-Sulfur Proteins / metabolism*
  • Mitochondria / genetics
  • Mitochondrial Diseases / genetics*
  • Mitochondrial Diseases / physiopathology
  • Mitochondrial Proteins / genetics
  • Mutation
  • Phenotype
  • Protein Conformation
  • Protein Multimerization
  • Stress, Physiological / genetics
  • Sulfur / metabolism

Substances

  • Caenorhabditis elegans Proteins
  • Carrier Proteins
  • Iron-Sulfur Proteins
  • Mitochondrial Proteins
  • NFU1 protein, human
  • Sulfur
  • Iron

Supplementary concepts

  • Multiple Mitochondrial Dysfunctions Syndrome

Grants and funding

This work was supported, in part, by the Intramural Research Program of the National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases (A.G.) and National Heart, Lung, and Blood Institute (M.N.S.). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.