Targeting of parvulin interactors by diazirine mediated cross-linking discloses a cellular role of human Par14/17 in actin polymerization.
Anna GoehringIrina MichinTina GerdesNina SchulzeMike BlueggelEdisa RehicFarnusch KaschaniMarkus KaiserPeter BayerPublished in: Biological chemistry (2021)
The peptidyl-prolyl cis/trans isomerases (PPIases) Parvulin 14 (Par14) and Parvulin 17 (Par17) result from alternative transcription initiation of the PIN4 gene. Whereas Par14 is present in all metazoan, Par17 is only expressed in Hominidae. Par14 resides mainly within the cellular nucleus, while Par17 is translocated into mitochondria. Using photo-affinity labeling, cross-linking and mass spectrometry (MS) we identified binding partners for both enzymes from HeLa lysates and disentangled their cellular roles. Par14 is involved in biogenesis of ribonucleoprotein (RNP)-complexes, RNA processing and DNA repair. Its elongated isoform Par17 participates in protein transport/translocation and in cytoskeleton organization. Nuclear magnetic resonance (NMR) spectroscopy reveals that Par17 binds to β-actin with its N-terminal region, while both parvulins initiate actin polymerization depending on their PPIase activity as monitored by fluorescence spectroscopy. The knockdown (KD) of Par17 in HCT116 cells results in a defect in cell motility and migration.
Keyphrases
- mass spectrometry
- magnetic resonance
- dna repair
- stem cells
- multiple sclerosis
- endothelial cells
- high resolution
- oxidative stress
- computed tomography
- single molecule
- transcription factor
- cell death
- pseudomonas aeruginosa
- signaling pathway
- ms ms
- liquid chromatography
- hepatitis c virus
- human immunodeficiency virus
- reactive oxygen species
- contrast enhanced
- gas chromatography
- nucleic acid