TY - JOUR
T1 - Pptc7 is an essential phosphatase for promoting mammalian mitochondrial metabolism and biogenesis
AU - Niemi, Natalie M.
AU - Wilson, Gary M.
AU - Overmyer, Katherine A.
AU - Vögtle, F. Nora
AU - Myketin, Lisa
AU - Lohman, Danielle C.
AU - Schueler, Kathryn L.
AU - Attie, Alan D.
AU - Meisinger, Chris
AU - Coon, Joshua J.
AU - Pagliarini, David J.
N1 - Funding Information:
We thank members of the Pagliarini laboratory for helpful discussions and Amy Lin for her assistance with figure generation. The authors thank the Genome Editing and Animal Modeling core at the University of Wisconsin Biotechnology Center, particularly Kathy Krentz and Dustin Rubinstein, for their design and creation of the CRISPR-Cas9 Pptc7 knockout model. We also thank the University of Wisconsin Biotechnology Center DNA Sequencing Facility for providing next generation sequencing (NGS) services, and the UWBC Bioinformatics Resource Center for the analysis of the NGS data. We thank Ben August and the UW Electron Microscope (EM) Facility for processing samples and for providing training and expertize in EM data acquisition and analysis. We thank Benjamin Des Soye and Michael Jewett for protocols and assistance with the cell-free protein synthesis (CFPS) experiments. We thank Elizabeth Craig and her laboratory for the cytochrome b2-(167)Δ19-DHFR plasmid, advice, and protocols on mitochondrial import assays. We thank Toshiya Endo for the gift of the GAL7::TIM50 strain used in the study. Research reported in this publication was supported by the National Institute of Diabetes and Digestive and Kidney Diseases and National Institute of General Medical Sciences of the National Institutes of Health under award numbers R01DK098672 (to D.J.P.), T32DK007665 (to N.M.N.), and P41GM108538 (to J.J.C. and D.J.P). The Genome Editing and Animal Modeling core at UW is supported by a University of Wisconsin Carbone Cancer Center Support Grant (P30CA014520). This work was further supported by a Morgridge Postdoctoral Research Fellowship (to K.A.O.), the Deutsche Forschungsgemeinschaft (ID 403222702 - SFB 1381) and the Excellence Initiative of the German Federal & State Governments (EXC 294 BIOSS and EXC 2189 CIBSS) (to C.M. and F.N.V.), and the Emmy-Noether Programm of the Deutsche Forschungsgemeinschaft (to F.N.V).
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Mitochondrial proteins are replete with phosphorylation, yet its functional relevance remains largely unclear. The presence of multiple resident mitochondrial phosphatases, however, suggests that protein dephosphorylation may be broadly important for calibrating mitochondrial activities. To explore this, we deleted the poorly characterized matrix phosphatase Pptc7 from mice using CRISPR-Cas9 technology. Strikingly, Pptc7−/− mice exhibit hypoketotic hypoglycemia, elevated acylcarnitines and serum lactate, and die soon after birth. Pptc7−/− tissues have markedly diminished mitochondrial size and protein content despite normal transcript levels, and aberrantly elevated phosphorylation on select mitochondrial proteins. Among these, we identify the protein translocase complex subunit Timm50 as a putative Pptc7 substrate whose phosphorylation reduces import activity. We further find that phosphorylation within or near the mitochondrial targeting sequences of multiple proteins could disrupt their import rates and matrix processing. Overall, our data define Pptc7 as a protein phosphatase essential for proper mitochondrial function and biogenesis during the extrauterine transition.
AB - Mitochondrial proteins are replete with phosphorylation, yet its functional relevance remains largely unclear. The presence of multiple resident mitochondrial phosphatases, however, suggests that protein dephosphorylation may be broadly important for calibrating mitochondrial activities. To explore this, we deleted the poorly characterized matrix phosphatase Pptc7 from mice using CRISPR-Cas9 technology. Strikingly, Pptc7−/− mice exhibit hypoketotic hypoglycemia, elevated acylcarnitines and serum lactate, and die soon after birth. Pptc7−/− tissues have markedly diminished mitochondrial size and protein content despite normal transcript levels, and aberrantly elevated phosphorylation on select mitochondrial proteins. Among these, we identify the protein translocase complex subunit Timm50 as a putative Pptc7 substrate whose phosphorylation reduces import activity. We further find that phosphorylation within or near the mitochondrial targeting sequences of multiple proteins could disrupt their import rates and matrix processing. Overall, our data define Pptc7 as a protein phosphatase essential for proper mitochondrial function and biogenesis during the extrauterine transition.
UR - http://www.scopus.com/inward/record.url?scp=85069460863&partnerID=8YFLogxK
U2 - 10.1038/s41467-019-11047-6
DO - 10.1038/s41467-019-11047-6
M3 - Article
C2 - 31324765
AN - SCOPUS:85069460863
SN - 2041-1723
VL - 10
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 3197
ER -