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A Targeted Mutation Identified through pKa Measurements Indicates a Postrecruitment Role for Fis1 in Yeast Mitochondrial Fission. J Biol Chem 2016 09 23;291(39):20329-44

Date

08/09/2016

Pubmed ID

27496949

Pubmed Central ID

PMC5034033

DOI

10.1074/jbc.M116.724005

Abstract

The tail-anchored protein Fis1 is implicated as a passive tether in yeast mitochondrial fission. We probed the functional role of Fis1 Glu-78, whose elevated side chain pKa suggests participation in protein interactions. Fis1 binds partners Mdv1 or Dnm1 tightly, but mutation E78A weakens Fis1 interaction with Mdv1, alters mitochondrial morphology, and abolishes fission in a growth assay. In fis1Δ rescue experiments, Fis1-E78A causes a novel localization pattern in which Dnm1 uniformly coats the mitochondria. By contrast, Fis1-E78A at lower expression levels recruits Dnm1 into mitochondrial punctate structures but fails to support normal fission. Thus, Fis1 makes multiple interactions that support Dnm1 puncta formation and may be essential after this step, supporting a revised model for assembly of the mitochondrial fission machinery. The insights gained by mutating a residue with a perturbed pKa suggest that side chain pKa values inferred from routine NMR sample pH optimization could provide useful leads for functional investigations.

Author List

Koppenol-Raab M, Harwig MC, Posey AE, Egner JM, MacKenzie KR, Hill RB

Author

Ronald Blake Hill PhD Professor in the Biochemistry department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Amino Acid Substitution
GTP Phosphohydrolases
Mitochondria
Mitochondrial Dynamics
Mitochondrial Proteins
Mutation, Missense
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
jenkins-FCD Prod-399 190a069c593fb5498b7fcd942f44b7bc9cdc7ea1