Investigation into the Structure and Function of l-Mgm1 in inner mitochondrial membrane fusion

Document Type

Poster

Campus where you would like to present

Ellensburg

Event Website

https://digitalcommons.cwu.edu/source

Start Date

18-5-2020

Abstract

Mitochondria are key components of the cell as they provide energy in the form of ATP through cellular respiration. A balance of fission and fusion maintains a dynamic, inter-connected mitochondrial network, which is important for overall function. When fusion is disrupted, the mitochondrial network becomes fragmented, mitochondrial genomes are lost, and respiration ceases. Mitochondrial inner membrane fusion is mediated by the GTP hydrolase Mgm1 (Mitochondrial Genome Maintenance 1) in yeast. Yeast make an excellent model system to study mitochondrial physiology since they are not dependent on respiration for growth. Mgm1 exists as a membrane embedded long isoform (l-Mgm1) and a soluble short isoform (s-Mgm1). Both l-Mgm1 and s-Mgm1 are required for inner membrane fusion, but the role of l-Mgm1 remains unclear. Our goal is to determine the regions of l-Mgm1 that are important for fusion activity. To do this, we employed a polymerase chain reaction (PCR) splicing by overlapping extension (SOE) technique to make versions of l-Mgm1 with regions removed. Once constructed, we will test the mutated l-Mgm1 proteins with yeast physiology assays to determine effects on mitochondrial fusion, mitochondrial genome maintenance, and respiration. Construction of the l-Mgm1 mutants has proven challenging. I am currently optimizing the PCR conditions to complete this phase of the project. In the meantime, I am becoming familiar with yeast culturing techniques and physiological assays.

Faculty Mentor(s)

Derek Ricketson

Department/Program

Chemistry

Additional Mentoring Department

https://cwu.studentopportunitycenter.com/2020/04/investigation-into-the-structure-and-function-of-l-mgm1-in-inner-mitochondrial-membrane-fusion/

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May 18th, 12:00 PM

Investigation into the Structure and Function of l-Mgm1 in inner mitochondrial membrane fusion

Ellensburg

Mitochondria are key components of the cell as they provide energy in the form of ATP through cellular respiration. A balance of fission and fusion maintains a dynamic, inter-connected mitochondrial network, which is important for overall function. When fusion is disrupted, the mitochondrial network becomes fragmented, mitochondrial genomes are lost, and respiration ceases. Mitochondrial inner membrane fusion is mediated by the GTP hydrolase Mgm1 (Mitochondrial Genome Maintenance 1) in yeast. Yeast make an excellent model system to study mitochondrial physiology since they are not dependent on respiration for growth. Mgm1 exists as a membrane embedded long isoform (l-Mgm1) and a soluble short isoform (s-Mgm1). Both l-Mgm1 and s-Mgm1 are required for inner membrane fusion, but the role of l-Mgm1 remains unclear. Our goal is to determine the regions of l-Mgm1 that are important for fusion activity. To do this, we employed a polymerase chain reaction (PCR) splicing by overlapping extension (SOE) technique to make versions of l-Mgm1 with regions removed. Once constructed, we will test the mutated l-Mgm1 proteins with yeast physiology assays to determine effects on mitochondrial fusion, mitochondrial genome maintenance, and respiration. Construction of the l-Mgm1 mutants has proven challenging. I am currently optimizing the PCR conditions to complete this phase of the project. In the meantime, I am becoming familiar with yeast culturing techniques and physiological assays.

https://digitalcommons.cwu.edu/source/2020/COTS/39