More than just a ticket canceller: the mitochondrial processing peptidase tailors complex precursor proteins at internal cleavage sites

The Mitochondrial processing peptidase (MPP) is well known for cleaving off N-terminal targeting signals from mitochondrial precursor proteins. Here we show that MPP also processes more complex precursors at internal cleavage sites, separating polyproteins into distinct functional enzymes. This function is conserved among eukaryotes.

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More than just a ticket canceller: The mitochondrial processing peptidase matures complex precursor proteins at internal cleavage sites

10.1101/2020.07.02.183996 ◽

2020 ◽

Author(s):

Jana Friedl ◽

Michael R. Knopp ◽

Carina Groh ◽

Eyal Paz ◽

Sven B. Gould ◽

...

Keyword(s):

Mitochondrial Matrix ◽

Cleavage Sites ◽

Arginine Biosynthesis ◽

Mitochondrial Processing Peptidase ◽

Precursor Proteins ◽

Internal Site ◽

Processing Peptidase ◽

Polyprotein Precursor ◽

Internal Processing ◽

Additional Role

AbstractMost mitochondrial proteins are synthesized in the cytosol as precursors that carry N-terminal presequences. After import into mitochondria, these targeting signals are cleaved off by the mitochondrial processing peptidase MPP, giving rise to shorter mature proteins. Using the mitochondrial tandem protein Arg5,6 as a model substrate, we demonstrate that MPP has an additional role in preprotein maturation, beyond the removal of presequences. Arg5,6 is synthesized as a polyprotein precursor that is imported into the mitochondrial matrix and subsequently separated into two distinct enzymes that function in arginine biogenesis. This internal processing is performed by MPP, which cleaves the Arg5,6 precursor both at its N-terminus and at an internal site between the Arg5 and Arg6 parts. The peculiar organization and biogenesis of Arg5,6 is conserved across fungi and might preserve the mode of co-translational subunit association of the arginine biosynthesis complex of the polycistronic arginine operon in prokaryotic mitochondrial ancestors. Putative MPP cleavage sites are also present at the junctions in other mitochondrial fusion proteins from fungi, plants and animals. Our data suggest that, in addition to its role as “ticket canceller” for the removal of presequences, MPP exhibits a second, widely conserved activity as internal processing peptidase for complex mitochondrial precursor proteins.

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Mitochondrial Processing Peptidase: Multiple-Site Recognition of Precursor Proteins

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.1999.1703 ◽

1999 ◽

Vol 265 (3) ◽

pp. 611-616 ◽

Cited By ~ 46

Author(s):

Akio Ito

Keyword(s):

Multiple Site ◽

Mitochondrial Processing Peptidase ◽

Precursor Proteins ◽

Processing Peptidase ◽

Site Recognition

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Glycine-rich Region of Mitochondrial Processing Peptidase α-Subunit Is Essential for Binding and Cleavage of the Precursor Proteins

Journal of Biological Chemistry ◽

10.1074/jbc.m003110200 ◽

2000 ◽

Vol 275 (44) ◽

pp. 34552-34556 ◽

Cited By ~ 36

Author(s):

Yumiko Nagao ◽

Sakae Kitada ◽

Katsuhiko Kojima ◽

Hidehiro Toh ◽

Satoru Kuhara ◽

...

Keyword(s):

Rich Region ◽

Α Subunit ◽

Mitochondrial Processing Peptidase ◽

Precursor Proteins ◽

Processing Peptidase

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Mitochondrial Processing Peptidase: Multiple-Site Recognition of Precursor Proteins

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.2000.2156 ◽

2000 ◽

Vol 268 (2) ◽

pp. 663

Author(s):

Akio Ito

Keyword(s):

Multiple Site ◽

Mitochondrial Processing Peptidase ◽

Precursor Proteins ◽

Processing Peptidase ◽

Site Recognition

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Structural requirement for recognition of the precursor proteins by the mitochondrial processing peptidase.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)31443-6 ◽

1994 ◽

Vol 269 (40) ◽

pp. 24673-24678

Author(s):

W.J. Ou ◽

T. Kumamoto ◽

K. Mihara ◽

S. Kitada ◽

T. Niidome ◽

...

Keyword(s):

Structural Requirement ◽

Mitochondrial Processing Peptidase ◽

Precursor Proteins ◽

Processing Peptidase

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Maturation of Mitochondrially Targeted Prx V Involves a Second Cleavage by Mitochondrial Intermediate Peptidase That Is Sensitive to Inhibition by H2O2

Antioxidants ◽

10.3390/antiox10030346 ◽

2021 ◽

Vol 10 (3) ◽

pp. 346

Author(s):

Juhyun Sim ◽

Jiyoung Park ◽

Hyun Ae Woo ◽

Sue Goo Rhee

Keyword(s):

Short Form ◽

Mitochondrial Matrix ◽

Mitochondrial Processing Peptidase ◽

Mouse Tissues ◽

N Terminus ◽

Mitochondrial Intermediate Peptidase ◽

Subsequent Degradation ◽

Processing Peptidase ◽

Mitochondria Targeting ◽

Over Time

Prx V mRNA contains two in-frame AUG codons, producing a long (L-Prx V) and short form of Prx V (S-Prx V), and mouse L-Prx V is expressed as a precursor protein containing a 49-amino acid N-terminal mitochondria targeting sequence. Here, we show that the N-terminal 41-residue sequence of L-Prx V is cleaved by mitochondrial processing peptidase (MPP) in the mitochondrial matrix to produce an intermediate Prx V (I-Prx V) with a destabilizing phenylalanine at its N-terminus, and further, that the next 8-residue sequence is cleaved by mitochondrial intermediate peptidase (MIP) to convert I-Prx V to a stabilized mature form that is identical to S-Prx V. Further, we show that when mitochondrial H2O2 levels are increased in HeLa cells using rotenone, in several mouse tissues by deleting Prx III, and in the adrenal gland by deleting Srx or by exposing mice to immobilized stress, I-Prx V accumulates transiently and mature S-Prx V levels decrease in mitochondria over time. These findings support the view that MIP is inhibited by H2O2, resulting in the accumulation and subsequent degradation of I-Prx V, identifying a role for redox mediated regulation of Prx V proteolytic maturation and expression in mitochondria.

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