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Protein interaction and phosphorylation of diaphanous 1 regulate mitochondria movement in H295R human adrenocortical cells
Author(s) -
Li Donghui,
Sewer Marion B
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.510.5
Subject(s) - rhoa , chemistry , mitochondrion , microbiology and biotechnology , phosphorylation , biology , signal transduction
Our lab has previously shown that mitochondrial movement regulates production of cortisol and adrenal androgen by adrenocorticotropin (ACTH) and dibutyryl cAMP (Bt 2 cAMP) stimulation. This movement is dependent on phosphorylation of RhoA and the RhoA effector protein diaphanous 1 (DIAPH1). DIAPH1 promotes the polymerization, and nucleation and branching of actin filaments. Based on these data, we hypothesized that ACTH/Bt 2 cAMP‐stimulated mitochondria movement requires the formation of a macromolecular complex containing DIAPH1. To test this hypothesis, we used mass spectrometry to identify proteins bound to DIAPH1. We found that DIAPH1 binds to several proteins, including microtubule‐associated protein 4, dynamin‐1, kinesin, β‐actin, and g‐actin. Further, we also identified that DIAPH1 is phosphorylated at multiple sites. Using time‐lapsed video microscopy, we found that overexpression of DIAPH1 wt ‐GFP but not mutants (DIAPH1 S1067D –GFPand DIAPH1 Y1097F ‐GFP) increases mitochondria movement. These results suggest that protein interaction and phosphorylation of DIAPH1 play a key role in the regulating the rate of mitochondria movement. This study is supported by MCB0818119 from the NSF.