Tether mutations that restore function and suppress pleiotropic phenotypes of the C. elegans isp-1(qm150) Rieske iron-sulfur protein.
Mitochondria play an important role in numerous diseases as well as normative aging. Severe reduction in mitochondrial function contributes to childhood disorders such as Leigh Syndrome, whereas mild disruption can extend the lifespan of model organisms. The Caenorhabditis elegans isp-1 gene encodes the Rieske iron-sulfur protein subunit of cytochrome c oxidoreductase (complex III of the electron transport chain). The partial loss of function allele, isp-1(qm150), leads to several pleiotropic phenotypes. To better understand the molecular mechanisms of ISP-1 function, we sought to identify genetic suppressors of the delayed development of isp-1(qm150) animals. Here we report a series of intragenic suppressors, all located within a highly conserved six amino acid tether region of ISP-1. These intragenic mutations suppress all of the evaluated isp-1(qm150) phenotypes, including developmental rate, pharyngeal pumping rate, brood size, body movement, activation of the mitochondrial unfolded protein response reporter, CO2 production, mitochondrial oxidative phosphorylation, and lifespan extension. Furthermore, analogous mutations show a similar effect when engineered into the budding yeast Rieske iron-sulfur protein Rip1, revealing remarkable conservation of the structure-function relationship of these residues across highly divergent species. The focus on a single subunit as causal both in generation and in suppression of diverse pleiotropic phenotypes points to a common underlying molecular mechanism, for which we propose a "spring-loaded" model. These observations provide insights into how gating and control processes influence the function of ISP-1 in mediating pleiotropic phenotypes including developmental rate, movement, sensitivity to stress, and longevity.
Department of Pathology, University of Washington, Seattle, WA 98195;
Gholamali
G
Jafari
Department of Pathology, University of Washington, Seattle, WA 98195;
Brian M
BM
Wasko
Department of Pathology, University of Washington, Seattle, WA 98195;
Ashley
A
Tonge
Department of Pathology, University of Washington, Seattle, WA 98195;
Nathan
N
Schurman
Department of Pathology, University of Washington, Seattle, WA 98195;
Cindy
C
Dong
Department of Pathology, University of Washington, Seattle, WA 98195;
Zhongyu
Z
Li
Department of Pathology, University of Washington, Seattle, WA 98195;
Rebecca
R
Peters
Center for Developmental Therapeutics, Seattle Children's Research Institute, Seattle, WA 98101;
Ernst-Bernhard
EB
Kayser
Department of Pathology, University of Washington, Seattle, WA 98195;
Jason N
JN
Pitt
Center for Developmental Therapeutics, Seattle Children's Research Institute, Seattle, WA 98101;
Phil G
PG
Morgan
Center for Developmental Therapeutics, Seattle Children's Research Institute, Seattle, WA 98101;
Margaret M
MM
Sedensky
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Antony R
AR
Crofts
Department of Pathology, University of Washington, Seattle, WA 98195; kaeber@uw.edu.
Matt
M
Kaeberlein
D000818:Animals; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D052284:Clutch Size / Q000235:genetics; D014450:Electron Transport Complex III / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D058685:Genetic Pleiotropy / Q000235:genetics*; D048788:Growth and Development / Q000235:genetics; D008136:Longevity / Q000235:genetics; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D009068:Movement / Q000502:physiology; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics; D028861:Nuclear Pore Complex Proteins / Q000235:genetics; D010641:Phenotype; D015202:Protein Engineering; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013312:Stress, Physiological / Q000235:genetics
D029742:Caenorhabditis elegans Proteins; C109212:NUP42 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D029701:Saccharomyces cerevisiae Proteins; C443102:Isp-1 protein, Caenorhabditis elegans; D014450:Electron Transport Complex III
Rip1; aging; complex III; isp-1; mitochondrial iron–sulfur protein