Yeast two-hybrid screening identifies binding partners of human Tom34 that have ATPase activity and form a complex with Tom34 in the cytosol.

In the accompany paper (Mukhopadhyay, A., Avramova, L. V. and Weiner, H., Arch. Biochem. Biophys.), it was shown that Tom34, a previously proposed putative translocase of the mitochondrial outer membrane, binds to the mature region of a precursor protein and appears to be a cytosol protein. Here Tom34 was used ...
as bait in a yeast two-hybrid screening to search for its potential binding partners. Two of the identified proteins were the ATPase-related valosin-containing protein (VCP) and the lysosomal H(+)-transporting ATPase member M (ATP6M). Tom34 was found primarily in the cytosol while VCP and ATP6M were found in the cytosol as well as in nonmitochondrial organelles. Tom34 formed a approximately 400-kDa complex with them in the cytosol. Tom34 was found to possess a weak ATPase activity that did not change when associated with VCP. The tetratricopeptide repeat (TPR) motif region of Tom34 (residue 201-256) was responsible for binding to the other proteins. Tom34 appears not to be a member of the mitochondrial outer membrane translocase family but might function as a chaperone-like protein during protein translocation.
Mesh Terms:
Adenosine Triphosphatases, Animals, Carrier Proteins, Cloning, Molecular, Cytosol, Electrophoresis, Gel, Two-Dimensional, Hela Cells, Humans, Hydrolysis, Isoelectric Focusing, Liver, Microscopy, Fluorescence, Mitochondria, Mitochondrial Membrane Transport Proteins, Protein Binding, Protein Structure, Tertiary, Protein Transport, Rats, Recombinant Proteins, Subcellular Fractions, Two-Hybrid System Techniques
Arch. Biochem. Biophys.
Date: Apr. 01, 2002
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