C ONCLUSIONS  AND   O UTLOOK

The mitochondrial inner membrane is morphologically dynamic. Various inner membrane proteins are known to contribute to the shape and structure of cristae membranes. F1FOATPase dimers contribute to a strong local curvature and thus the angular shape of the cristae apex (Strauss et al., 2008; Seelert and Dencher, 2011).

Mutants that lack the Atp20 or Atp21 dimerization factors exemplify the importance of F1FOATPase dimerization with respect to cristae morphology. These mutants clearly exhibit mutant inner mitochondrial organization that characterized by concentric

‘onion-like’ structures (Paumard et al., 2002b; Arselin et al., 2003; Arselin et al., 2004). Moreover, Mitofilins (Fcj1 in yeast) has been previously shown to be an important determinant of cristae junction formation by acting in an antagonistic manner in inner membrane curvature with respect to F1FOATPase dimers (Rabl et al., 2009). Whereas this study did not result the identification of novel F1FOATPase dimerization factors, Mio10/MINOS1 are identified as novel interaction partners of Fcj1/Mitofilin. This study also provided evidence for the MINOS complex playing a critical role in inner membrane architecture. The role of Mio10 in cristae morphology in yeast was shown by the analysis of MIO10 deletion mutants in comparison to other strains. The results of this study is paralleled and verified by three other studies, which have in addition identified four other components of the MINOS complex (Hoppins et

DISCUSSION

Results from this study and other publications support the notion that MINOS is a multifunctional complex (Figure 36). Beside the obvious role in cristae morphology, the MINOS may play a role in mitochondrial protein import by its interaction with SAM, TOM, and MIA components (Xie et al., 2007; von der Malsburg et al., 2011; Harner et al., 2011). By its interaction with Ugo1, the MINOS complex plays a role in fusion and mitochondrial network dynamics (Harner et al., 2011). Moreover, it may have a role in mitochondrial inheritance via its components, Aim5, Aim13, and Aim37 (Hess et al., 2009). MINOS interaction with cytoskeleton proteins (FAM82B and Tubulin) may indicate its role in mitochondrial trafficking.

Furthermore it may play a role in ERMES and ER mitochondrial dynamics via its interaction with ER resident proteins (TANGO1). This notion is further supported by a strong genetic interaction of MINOS and ERMES genes (Hoppins et al., 2011).

Besides its link to apoptosis via Mitofilin interaction with OPA1 (Darshi et al., 2011), the MINOS complex may also play role in cancer. Indeed, MINOS1 proteins levels are elevated in prostate cancer cells (Lin et al., 2005). In addition, overexpression of CHCHD6 in human cancer cells enhances resistance to genotoxic anticancer drugs, whereas its knockdown increases its chemosensitivity (An et al., 2012). Other than its proven role in inner membrane architecture, any role MINOS might have in other mitochondrial and non mitochondrial processes remains speculative and further investigations are necessary.

DISCUSSION

Figure 36. MINOS is a multifunctional protein. Besides the MINOS role in cristae morphology, results from various sources indicate potential MINOS interactions with ERMES (via SAM and TANGO1), mitochondrial trafficking (Tubulin), mitochondrial import (TOM, SAM, Mia40), mitochondrial fusion/fission (Ugo1), and mitochondrial inheritance (AIM proteins of the MINOS complex).

Besides the characterization of novel roles MINOS may have in mitochondria and other cellular organelles, information is still required with respect to the structure and exact function of MINOS in inner membrane organization. For instance, the exact composition of the MINOS complex in higher eukaryotes remains an open question.

Still, it may be postulated from available data that MINOS in higher eukaryotes contains at least MINOS1, Mitofilin, CHCHD3 and CHCHD6. It should also be considered that the composition of the MINOS complex might vary across the species divide. A homolog of Aim5 and Aim13 is not found in higher eukaryotes whereas a homolog for CHCHD3 is not found in yeast (Herrmann, 2011). The MINOS complex may require at least Mio10/MINOS1 and Fcj1/Mitofilin, whereas other components

DISCUSSION

RNAi in human mitochondria and whether the role of MINOS1 in the MINOS complex is as essential as the yeast Mio10.

More evidence is still required to understand how the MINOS complex and its components exert their function in cristae biogenesis. It should be noted that most of the of evidence related to the roles of these inner membrane proteins have been deduced from tomographies of mutants, whether yeast or human cultured cells. These tomographies represent single-point illustrations of the mutations effects but provide limited insight onto the dynamics governing how cristae membranes are rendered

‘online-like’, ‘stacked’, or ‘vesicle-like’. This further highlights the extent to which cristae propagation, maintenance, and dynamics remain to be studied.

Analysis of the MINOS mutants indicted that that MINOS complex affects cristae junction opening diameters (Rabl et al., 2009; von der Malsburg et al., 2011;

Hoppins et al., 2011). It also plays a role in maintenance of cristae tips, exemplified by the lack of cristae tips in various MINOS mutants. In the absence of MINOS components, the cristae membranes appear detached from the IBM and form stacks that are linked together with the F1FOATPase (Rabl et al., 2009; von der Malsburg et al., 2011). MINOS might thus have a role in cristae biogenesis: cristae tip formation, cristae membrane curvature, junction maintenance; and cristae membrane propagation (Figure 37).

DISCUSSION

Figure 37. Overview of MINOS roles in cristae biogenesis. A) By inducing positive membrane curvature, MINOS leads to invagination of the inner membrane and eventual cristae tip formation. In the absence of MINOS, cristae tips are not found. B) The large molecular size of MINOS might regulate cristae junction width. In the absence of some MINOS components, cristae membranes appear to have narrower cristae junctions. C) MINOS might have a role in the regulation cristae fusion. In the absence of MINOS, fission of cristae membrane is active whereas fusion discontinues. This may lead to an increase in cristae vesicle, which are then stacked by F1FOATPase dimers.

MINOS may affect cristae junction due its large molecular size and by its interaction with OPA-1. OPA-1 controls cristae opening size by forming a scaffold at the opening that prevents premature release of cytochrome c under non-apoptotic conditions (Frezza et al., 2006). The role of MINOS in cristae biogenesis may be

DISCUSSION

enough to explain the observed stacked cristae inner membranes of MINOS mutants.

In this case, a cristae fusion/fission model may be applied. This model proposes that cristae formation is dynamic, in which cristae membrane vesicles may continuously fuse from one site of the inner membrane and fuse to another (Zick et al., 2009). The notion that cristae propagation is dynamic is supported by the observation of certain reversible cristae morphologies (Mannella et al., 2001). Stacked membranes in MINOS mutants may thus be due to continuous cristae fission but a lack of fusion. The MINOS complex may play a role in the equilibrium between cristae fusion and fission.

Information is still lacking to explain a variation of phenotypes within any single MINOS mutants (wildtype, onion-like, or intermediate). Thus, it may seem that other factors may be involved with respect to MINOS and cristae morphology. Further analysis such as the analysis of multiple deletions or knockouts of MINOS components may shine light on the exact role of MINOS. Moreover, high-resolution immunoelectron microscopy analysis of mitochondrial inner membrane in the presence or absence of MINOS components may provide novel insight into the mechanism of the complex.

MINOS is not only an inner membrane organizer that forms a scaffold connecting the inner and outer mitochondrial membranes, but may feature in various other mitochondrial and extra-mitochondrial functions. Many of these functions are yet to be examined and further studies are required. Various outstanding questions remain with respect to the exact role of the MINOS complex. Nonetheless, the discovery of new MINOS components here may provide new insight and direction for future studies.

REFERENCES