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. 2007 Jul 2;178(1):71–84. doi: 10.1083/jcb.200611064

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Copyright © 2007, The Rockefeller University Press

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Figure 1. — Mutations of the MARCH5 RING domain induce abnormal morphology of mitochondria. MARCH5 structure and the sequence homology of RINGv domains of various E3 ubiquitin ligases. The conserved histidine 43 (red) and cysteines 65 and 68 (blue) that are predicted to affect MARCH5 RING finger domain activity were mutated resulting in H43W and C65S,C68S mutants (A). Mitochondrial morphology in HeLa cells expressing wild type MARCH5-YFP, (green; B), MARCH5^H43W-YFP (green; C and D), and MARCH5^C65S,C68S-YFP (green; E) was analyzed by confocal microscopy. Mitochondria (red) were revealed by staining for cytochrome c. Number of cells with control-like tubular mitochondria, abnormally connected mitochondria, and with mitochondria predominantly localizing in perinuclear regions were scored (F). Data represent the average ± SD of four transfections per condition with 100 cells counted each time.