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Identification of New OPA1 Cleavage Site Reveals that Short Isoforms Regulate Mitochondrial Fusion

Citation

Wang, Ruohan (2021) Identification of New OPA1 Cleavage Site Reveals that Short Isoforms Regulate Mitochondrial Fusion. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/8nk6-1502. https://resolver.caltech.edu/CaltechTHESIS:11182020-224422123

Abstract

OPA1 is a 120kDa large GTPase belonging to the dynamin superfamily. It is the only known mitochondrial inner membrane fusion protein, mediating fusion of the mitochondrial inner membranes following outer membrane fusion. Additionally, OPA1 also regulates cristae morphology and maintains respiratory chain function.

OPA1 has two forms—inner-membrane-anchored long forms (l-OPA1) and cleaved inter-membrane-space only short forms (s-OPA1). L-OPA1 are proteolytically processed by two mitochondrial proteases—OMA1 and YME1L, acting at cleavage sites S1 and S2 respectively, to produce s-OPA1.

In both mice and human, half of the mRNA splice forms of Opa1 are constitutively processed post translation to yield exclusively s-OPA1. However, the specific function of s-OPA1 in mitochondrial fusion has been debated—under basal conditions, s-OPA1 are needed to maintain optimal fusion activity, but in certain stress conditions, s-OPA1 is dispensable for fusion. By constructing cells in which the Opa1 locus no longer produces transcripts with S2 cleavage sites using CRISPR-Cas9, we generated a simplified system to identify the novel YME1L-dependent site S3 that mediates constitutive and complete cleavage of OPA1. We found that S3 site locates within the C-terminal leucine string of Opa1 exon4b, slightly upstream of the well-established S1. We show that mitochondrial morphology is highly sensitive to the ratio of l-OPA1 to s-OPA1, indicating that s-OPA1 fine tunes mitochondrial fusion.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:Mitochondria, inner membrane fusion, dynamics, Opa1, protease cleavage
Degree Grantor:California Institute of Technology
Division:Biology and Biological Engineering
Major Option:Biology
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Chan, David C.
Thesis Committee:
  • Bronner, Marianne E. (chair)
  • Voorhees, Rebecca M.
  • Aravin, Alexei A.
  • van der Bliek, Alexander M.
Defense Date:28 October 2020
Record Number:CaltechTHESIS:11182020-224422123
Persistent URL:https://resolver.caltech.edu/CaltechTHESIS:11182020-224422123
DOI:10.7907/8nk6-1502
Related URLs:
URLURL TypeDescription
https://doi.org/10.1091/mbc.E20-09-0605DOIArticle Adopted for Chapter 2
ORCID:
AuthorORCID
Wang, Ruohan0000-0001-8361-4005
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:14002
Collection:CaltechTHESIS
Deposited By: Ruohan Wang
Deposited On:05 Jan 2021 19:40
Last Modified:12 Jan 2021 18:33

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