A designed point mutant in Fis1 disrupts dimerization and mitochondrial fission.

Jonathan P B Lees, Cara Marie Manlandro, Lora K Picton, Alexandra Z Ebie Tan, Salvador Casares, John M Flanagan, Karen G Fleming, R Blake Hill

Journal of molecular biology 2012 Oct 19

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Mitochondrial and peroxisomal fission are essential processes with defects resulting in cardiomyopathy and neonatal lethality. Central to organelle fission is Fis1, a monomeric tetratricopeptide repeat (TPR)-like protein whose role in assembly of the fission machinery remains obscure. Two nonfunctional, Saccharomyces cerevisiae Fis1 mutants (L80P or E78D/I85T/Y88H) were previously identified in genetic screens. Here, we find that these two variants in the cytosolic domain of Fis1 (Fis1ΔTM) are unexpectedly dimeric. A truncation variant of Fis1ΔTM that lacks an N-terminal regulatory domain is also found to be dimeric. The ability to dimerize is a property innate to the native Fis1ΔTM amino acid sequence as we find this domain is dimeric after transient exposure to elevated temperature or chemical denaturants and is kinetically trapped at room temperature. This is the first demonstration of a specific self-association in solution for the Fis1 cytoplasmic domain. We propose a three-dimensional domain-swapped model for dimerization that is validated by a designed mutation, A72P, which potently disrupts dimerization of wild-type Fis1. A72P also disrupts dimerization of nonfunctional variants, indicating a common structural basis for dimerization. The obligate monomer variant A72P, like the dimer-promoting variants, is nonfunctional in fission, consistent with a model in which Fis1 activity depends on its ability to interconvert between monomer and dimer species. These studies suggest a new functionally important manner in which TPR-containing proteins may reversibly self-associate. Copyright © 2012 Elsevier Ltd. All rights reserved.

Citation

Jonathan P B Lees, Cara Marie Manlandro, Lora K Picton, Alexandra Z Ebie Tan, Salvador Casares, John M Flanagan, Karen G Fleming, R Blake Hill. A designed point mutant in Fis1 disrupts dimerization and mitochondrial fission. Journal of molecular biology. 2012 Oct 19;423(2):143-58