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Connection of Protein Transport and Organelle Contact Sites in Mitochondria rights and content


Mitochondria form contact sites to other cellular organelles.

Mitochondrial contact sites to the endoplasmic reticulum mediate lipid transfer.

Mitochondrial protein translocases are linked to organelle contact sites.


Mitochondrial biogenesis and function depend on the intensive exchange of molecules with other cellular compartments. The

mitochondrial outer membrane plays a central role in this communication process. It is equipped with a number of specific protein

machineries that enable the transport of proteins and metabolites. Furthermore, the outer membrane forms molecular contact sites

with other cell organelles like the endoplasmic reticulum (ER), thus integrating mitochondrial function in cellular physiology. The

best-studied mitochondrial organelle contact site, the ER–mitochondria encounter structure (ERMES) has been linked to many vital

processes including mitochondrial division, inheritance, mitophagy, and phospholipid transport. Strikingly, ER–mitochondria contact

sites are closely connected to outer membrane protein translocases. The translocase of the outer mitochondrial membrane (TOM)

represents the general mitochondrial entry gate for precursor proteins that are synthesized on cytosolic ribosomes. The outer

membrane also harbors the sorting and assembly machinery (SAM) that mediates membrane insertion of β-barrel proteins. Both of

these essential protein translocases are functionally linked to ER–mitochondria contact sites. First, the SAM complex associates

with an ERMES core component to promote assembly of the TOM complex. Second, several TOM components have been co-opted

as ER–mitochondria tethers. We propose that protein import and organelle contact sites are linked to coordinate processes

important for mitochondrial biogenesis.

Graphical Abstract
Crystal structure of Mdm12 and combinatorial reconstitution of Mdm12/Mmm1 ERMES complexes for structural studies rights and content


We solved a 4.1 Å resolution crystal structure of Mdm12, the soluble subunit of ERMES.

The two Mdm12 apo monomers differ in the conformation of their N-terminal β-strands.

We reconstituted and crystallized two distinct Mdm12/Mmm1 complexes.

SAXS analysis of the complexes agrees with our previous electron microscopy data.

The structure suggests a head-to-tail association in the Mdm12/Mmm1 complex.


Membrane contact sites between organelles serve as molecular hubs for the exchange of metabolites and signals. In yeast, the

Endoplasmic Reticulum – Mitochondrion Encounter Structure (ERMES) tethers these two organelles likely to facilitate the non-

vesicular exchange of essential phospholipids. Present in Fungi and Amoebas but not in Metazoans, ERMES is composed of five

distinct subunits; among those, Mdm12, Mmm1 and Mdm34 each contain an SMP domain functioning as a lipid transfer module. We

previously showed that the SMP domains of Mdm12 and Mmm1 form a hetero-tetramer. Here we describe our strategy to diversify

the number of Mdm12/Mmm1 complexes suited for structural studies. We use sequence analysis of orthologues combined to

protein engineering of disordered regions to guide the design of protein constructs and expand the repertoire of Mdm12/Mmm1

complexes more likely to crystallize. Using this combinatorial approach we report crystals of Mdm12/Mmm1 ERMES complexes

currently diffracting to 4.5 Å resolution and a new structure of Mdm12 solved at 4.1 Å resolution. Our structure reveals a monomeric

form of Mdm12 with a conformationally dynamic N-terminal β-strand; it differs from a previously reported homodimeric structure

where the N-terminal β strands where swapped to promote dimerization. Based on our electron microscopy data, we propose a

refined pseudo-atomic model of the Mdm12/Mmm1 complex that agrees with our crystallographic and small-angle X-ray scattering

(SAXS) solution data.



Membrane contact sites

Mdm12/Mmm1 complex reconstitution

SMP domain

Crystal structure