Mitochondria are complex organelles with a highly dynamic distribution and internal organization. Here, we demonstrate that mitofilin, a previously identified mitochondrial protein of unknown function, controls mitochondrial cristae morphology. Mitofilin is enriched in the narrow space between the inner boundary and the outer membranes, where it forms a homotypic interaction and assembles into a large multimeric protein complex. Down-regulation of mitofilin in HeLa cells by using specific small interfering RNA lead to decreased cellular proliferation and increased apoptosis, suggesting abnormal mitochondrial function. Although gross mitochondrial fission and fusion seemed normal, ultrastructural studies revealed disorganized mitochondrial inner membrane. Inner membranes failed to form tubular or vesicular cristae and showed as closely packed stacks of membrane sheets that fused intermittently, resulting in a complex maze of membranous network. Electron microscopic tomography estimated a substantial increase in inner:outer membrane ratio, whereas no cristae junctions were detected. In addition, mitochondria subsequently exhibited increased reactive oxygen species production and membrane potential. Although metabolic flux increased due to mitofilin deficiency, mitochondrial oxidative phosphorylation was not increased accordingly. We propose that mitofilin is a critical organizer of the mitochondrial cristae morphology and thus indispensable for normal mitochondrial function. INTRODUCTIONMitochondria are the center of cellular energy production and essential metabolic reactions. As double membranebound organelles, mitochondria from different species, tissues, and metabolic states are highly polymorphic in nature yet exhibit common structural features. The ultrastructural variations in mitochondrial architecture occur mainly due to the differences in the amount and shape of cristae, which derive from the infolded inner membrane in which protein complexes of oxidative phosphorylation and intermediate metabolism are embedded. Abundant cristae are found in mitochondria from tissues where energy demand is high. For example, mitochondria with densely packed cristae are observed in the flight muscle of the dragonfly, whereas the liver of a winter-starved frog displays mitochondria with sparse cristae (Ghadially, 1997). Furthermore, the inner membranes of isolated mitochondria undergo characteristic morphological changes in response to the metabolic state (Hackenbrock, 1966). Although little is known about the molecular mechanisms regulating cristae biogenesis and architecture, recent studies have implicated proteins resident in both the outer and inner membrane to have roles in this process.The mitochondrial fission and fusion machinery plays an essential role in the dynamics, division, distribution, and morphology of the organelle (Yaffe, 1999;Jensen et al., 2000;Griparic and van der Bliek, 2001;Shaw and Nunnari, 2002). Three evolutionarily conserved large GTPases, Dnm1/ Drp1/Dlp1, Fzo1/mitofusin, and Mgm1/MspI/OPA1, are...
The inner membrane system of mitochondria is known to consist of two contiguous but distinct membranes: the inner boundary membrane, which apposes the outer membrane, and the cristal membrane, which forms tubules or lamellae in the interior. Using immunolabeling and transmission electron microscopy of bovine heart tissue, we have calculated that around 94% of both Complex III of the respiratory chain and the ATP synthase are located in the cristal membrane, and only around 6% of either is in the inner boundary membrane. When accounting for the topographical ratio of cristal membrane versus inner boundary membrane, we ¢nd that both complexes exist at a 2.2^2.6-fold higher concentration in the cristal membrane. The residual protein in the inner boundary membrane may be newly assembled complexes destined for cristal membranes. Our results argue for restricted di¡usion of complexes through the cristal junctions and indicate that the mitochondrial cristae comprise a regulated submitochondrial compartment specialized for ATP production.
Depletion of mitochondrial DNA (mtDNA) causes defects in respiratory activity and energy production. Recent studies have shown mitochondria to exist primarily as reticular networks, having tubular cristae. Using fluorescence microscopy and transmission electron microscopy, we have examined mitochondrial morphology and interior structure in wildtype and mtDNA-depleted b b0 human fibroblasts and 143B osteosarcoma cell lines. MtDNA depletion results in compromise of the mitochondrial continuum and causes a reduction in amount of cristal membranes, often prompting the remaining cristae to adopt a circular appearance in the mitochondrial interior. These changes emphasize the tight relationship between mitochondrial structure and function.z 2000 Federation of European Biochemical Societies.
DNA methylation, heterochromatin protein 1 (HP1), histone H3 lysine 9 (H3K9) methylation, histone deacetylation, and highly repeated sequences are prototypical heterochromatic features, but their interrelationships are not fully understood. Prior work showed that H3K9 methylation directs DNA methylation and histone deacetylation via HP1 in Neurospora crassa and that the histone deacetylase complex HCHC is required for proper DNA methylation. The complex consists of the chromodomain proteins HP1 and chromodomain protein 2 (CDP-2), the histone deacetylase HDA-1, and the AT-hook motif protein CDP-2/HDA-1–associated protein (CHAP). We show that the complex is required for proper chromosome segregation, dissect its function, and characterize interactions among its components. Our analyses revealed the existence of an HP1-based DNA methylation pathway independent of its chromodomain. The pathway partially depends on CHAP but not on the CDP-2 chromodomain. CDP-2 serves as a bridge between the recognition of H3K9 trimethylation (H3K9me3) by HP1 and the histone deacetylase activity of HDA-1. CHAP is also critical for HDA-1 localization to heterochromatin. Specifically, the CHAP zinc finger interacts directly with the HDA-1 argonaute-binding protein 2 (Arb2) domain, and the CHAP AT-hook motifs recognize heterochromatic regions by binding to AT-rich DNA. Our data shed light on the interrelationships among the prototypical heterochromatic features and support a model in which dual recognition by the HP1 chromodomain and the CHAP AT-hooks are required for proper heterochromatin formation.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.