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Mimicking human Drp1 disease-causing mutations in yeast Dnm1 reveals altered mitochondrial dynamics
Riddhi Banerjee1, Abhishek Kumar2, Priyadarshi Satpati2
1Organelle Biology and Cellular Ageing Lab, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati 781039, Assam, India.
Abstract:
The dynamin-related protein 1 (Drp1) and its homologs in various eukaryotes are essential to maintain mitochondrial morphology and regulate mitochondrial division. Several mutations in different domains of Drp1 have been reported, which result in debilitating conditions. Four such disease-causing mutations of the middle domain of Drp1 were mimicked in the yeast dynamin-related GTPase (Dnm1) and were characterized in this study. Mitochondrial morphology and protein function were observed to be altered to a variable extent in cells expressing the mutated variants of Dnm1. Several aspects related to the protein such as punctate formation, localization to mitochondria, dynamic behavior and structure were analyzed by microscopy, biochemical studies and molecular dynamics simulations. Significant effects on the protein structure and function were observed in cells expressing A430D and G397D mutations. Overall, our data provide insight into the molecular and cellular alterations resulting from middle domain mutations in Dnm1.
Insights
Middle domain mutations in dynamin-related GTPase (Dnm1) disrupt mitochondrial morphology and protein function. Specific mutations, A430D and G397D, significantly altered Dnm1 structure and cellular behavior.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Dynamin-related protein 1 (Drp1) is crucial for mitochondrial dynamics and morphology.
- Mutations in Drp1 are linked to severe human diseases.
- The middle domain of Drp1 is a key regulatory region.
Purpose of the Study:
- To investigate the functional and structural consequences of disease-associated mutations in the middle domain of yeast dynamin-related GTPase (Dnm1).
- To analyze how these mutations affect mitochondrial morphology, protein localization, and dynamic behavior.
Main Methods:
- Expression of four disease-mimicking Dnm1 middle domain mutations in yeast.
- Microscopy to assess mitochondrial morphology and protein localization.
- Biochemical assays to evaluate protein function.
- Molecular dynamics simulations to analyze structural changes.
Main Results:
- Mutated Dnm1 variants exhibited altered mitochondrial morphology and protein function to varying degrees.
- A430D and G397D mutations caused significant disruptions in Dnm1 structure and cellular activity.
- Analysis revealed changes in punctate formation, mitochondrial localization, and dynamic behavior.
Conclusions:
- Middle domain mutations in Dnm1 can lead to significant molecular and cellular defects.
- Specific mutations (A430D, G397D) have pronounced effects on Dnm1 structure and function.
- This study provides insights into the pathogenic mechanisms of Drp1-related disorders.
