Submission Title

Identification of Protein Interactions for the Mitochondrial Transcription Factor TFAM and Mutants

Subject Area

Biochemistry

Description

Mitochondria are key organelles in eukaryotic cells for their role in metabolism and other biosynthetic pathways. They play a key role in the production of ATP via oxidative phosphorylation (OXPHOS). Over eighty proteins make up the various OXPHOS complexes, several of which are encoded by the mitochondrial genome (mtDNA). Within mitochondria the processes of transcription and translation take place to generate these important OXPHOS subunit proteins. During the process of mitochondrial transcription, the transcription factor TFAM (transcription factor A, mitochondria) is important in promoter regulation. TFAM itself is a multifunctional mitochondrial protein in that it binds both specific and nonspecific mtDNA sequences. At promoter sequences the protein causes U-turn in the DNA, while less dramatic bending takes places when bound nonspecifically. Our aim is to identify novel interacting proteins with TFAM using a yeast-two-hybrid model. Additionally, we are interested in determining if there are changes in these protein interacting partners in mutant forms of TFAM. Specially, two point mutations in the TFAM gene have been linked to late onset Alzheimer’s disease, S12T and P178L. Biochemical and genetic techniques are being used to identify and characterize changes in protein interaction partners as a result of these mutations.

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Identification of Protein Interactions for the Mitochondrial Transcription Factor TFAM and Mutants

Mitochondria are key organelles in eukaryotic cells for their role in metabolism and other biosynthetic pathways. They play a key role in the production of ATP via oxidative phosphorylation (OXPHOS). Over eighty proteins make up the various OXPHOS complexes, several of which are encoded by the mitochondrial genome (mtDNA). Within mitochondria the processes of transcription and translation take place to generate these important OXPHOS subunit proteins. During the process of mitochondrial transcription, the transcription factor TFAM (transcription factor A, mitochondria) is important in promoter regulation. TFAM itself is a multifunctional mitochondrial protein in that it binds both specific and nonspecific mtDNA sequences. At promoter sequences the protein causes U-turn in the DNA, while less dramatic bending takes places when bound nonspecifically. Our aim is to identify novel interacting proteins with TFAM using a yeast-two-hybrid model. Additionally, we are interested in determining if there are changes in these protein interacting partners in mutant forms of TFAM. Specially, two point mutations in the TFAM gene have been linked to late onset Alzheimer’s disease, S12T and P178L. Biochemical and genetic techniques are being used to identify and characterize changes in protein interaction partners as a result of these mutations.