SUMMARY
Progesterone Receptor Membrane Component 1 (PGRMC1) is expressed in many cancer cells, where it is associated with detrimental patient outcomes. It contains phosphorylated tyrosines which evolutionarily preceded deuterostome gastrulation and tissue differentiation mechanisms. Here, we demonstrate that manipulating PGRMC1 phosphorylation status in MIA PaCa-2 (MP) cells imposes broad pleiotropic effects. Relative to parental cells over-expressing hemagglutinin-tagged wild-type (WT) PGRMC1-HA, cells expressing a PGRMC1-HA-S57A/S181A double mutant (DM) exhibited reduced levels of proteins involved in energy metabolism and mitochondrial function, and altered glucose metabolism suggesting modulation of the Warburg effect. This was associated with increased PI3K/Akt activity, altered cell shape, actin cytoskeleton, motility, and mitochondrial properties. An S57A/Y180F/S181A triple mutant (TM) indicated the involvement of Y180 in PI3K/Akt activation. Mutation of Y180F strongly attenuated mouse xenograft tumor growth. An accompanying paper demonstrates altered metabolism, mutation incidence, and epigenetic status in these cells, indicating that PGRMC1 phosphorylation strongly influences cancer biology.
Footnotes
↵2 Co-first author.
Manuscript was reformatted, with re-arrangement of some figure panels and editing of text, and shortening of title. The data and conclusions are unaltered.
- LIST OF ABBREVIATIONS
- adjP
- Benjamini-Hochberg adjusted p-value
- avFF
- average mitochondrial form factor
- CK2
- Casein kinase 2
- DM
- hemagglutinin-tagged PGRMC1 S57A/S181 double mutant
- EMT
- epithelial-mesenchymal transition
- ERR1
- estrogen receptor related 1
- FF
- mitochondrial form factor
- hPSCs
- human pluripotent stem cells
- IDA
- information dependent acquisition
- LDLR
- low density lipoprotein receptor
- MAT
- mesenchymal-amoeboid transition
- MP
- MIA PaCa-2 pancreatic cancer
- MS/MS
- tandem mass spectrometry
- MTT
- 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide
- NpFR2
- Naphthalimide-flavin redox sensor 2
- P4
- progesterone
- PAQR7
- progestin and adipoQ receptor 7
- PGRMC1
- Progesterone Receptor Membrane Component 1
- PGRMC2
- Progesterone Receptor Membrane Component 2
- ROCK
- Rho kinase
- ROCKI
- Rho kinase inhibitor
- RPPA
- Reverse Phase Protein Array
- S2R
- Sigma-2 receptor
- SH2
- Src homology 2
- SH3
- Src homology 3
- SWATH-MS
- Sequential Window Acquisition of all Theoretical Mass Spectrometry
- TM
- hemagglutinin-tagged PGRMC1 S57A/Y180F/S181 triple mutant
- TRiC
- T-complex protein-1 ring complex
- WT
- hemagglutinin-tagged PGRMC1 wild type
- Δψm
- mitochondrial membrane potential