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Fig. 6. DLP1 binding to hFis1, but not the DLP1-GTPase activity, is necessary for mitochondrial swelling induced by overexpression of 
1-deleted hFis1. (A) Leucine-to-proline mutants (L42P, L58P, L77P, L91P, and L110P) in Myc-hFis1[32-152] were expressed in Clone 9 cells and mitochondrial morphologies were analyzed. The TPR mutants (L42P, L58P, L77P, and L91P) in Myc-hFis1[32-152] significantly decreased the mitochondrial swelling phenotype of Myc-hFis1[32-152] although L42P was less effective, indicating that DLP1 binding to the hFis1 TPR is necessary for mitochondrial swelling. The L110P mutation had no effect on mitochondrial swelling. (B) Co-expression of the GTPase-defective DLP1-K38A with Myc-hFis1[32-152] did not reduce the mitochondrial swelling, but mitochondrial fragmentation by Myc-hFis1-FL was greatly decreased by DLP1-K38A, indicating that GTP hydrolysis by DLP1 is required for mitochondrial fission but not for the swelling. (C) Although mitochondrial swelling via 1-deletion (Myc-hFis1[32-152]) was reduced by the Bcl-2 co-expression, Bcl-2 had no effect on mitochondrial fragmentation induced by Myc-hFis1-FL, indicating that Bcl-2 does not directly participate in DLP1/hFis1-mediated mitochondrial fission.
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