Targeting of LRRC59 to the Endoplasmic Reticulum and the Inner Nuclear Membrane
Abstract
:1. Introduction
2. Results
2.1. LRRC59 can be Post-Translationally Inserted into Microsomal Membranes
2.2. Membrane Insertion of LRRC59 Does Not Require the TRC40 System
2.3. LRRC59 Localizes to the INM
2.4. Importin β Is Not Required for Localization of LRRC59 at the INM
2.5. Nuclear Accumulation of LRRC59 Lacking the TMD
2.6. The Size of the Cytoplasmic Domain Controls INM Localization of LRRC59
3. Discussion
3.1. Post-Translational Targeting of LRRC59 to ER-Membranes
3.2. Targeting of LRRC59 to the INM
4. Materials and Methods
4.1. Plasmids and Constructs
4.2. Cell Culture, Plasmid- and siRNA- Transfection and Immunofluorescence Microscopy
4.3. SDS-PAGE, Ponceau S Staining and Western Blotting
4.4. Microsome Integration Assay
4.5. Rapamycin Induced Dimerization Assay
5. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
Abbreviations
CAML | Ca2+-modulating cyclophilin ligand |
CIP2A | cancerous inhibitor of PP2A |
cNLS | classical nuclear localization signal |
ER | endoplasmic reticulum |
EK-RM | EDAT and high salt treated rough microsomes |
FKBP12 | 12 kDa FK506 binding protein |
FG repeats | phenylalanine-glycine repeats |
FRB | FKBP rapamycin binding domain |
INM | inner nuclear membrane |
LBR | Lamin B receptor |
LRRC59 | leucine-rich repeat-containing protein 59 |
NES | nuclear export sequence |
NLS | nuclear localization signal |
NPC | nuclear pore complex |
ONM | outer nuclear membrane |
PNGase F | Peptide-N-glycosidase F |
RM | rough microsomes |
T-RM | trypsin-treated rough microsomes |
TA | tail-anchored |
TMD | Transmembrane domain |
TRC40 | ATPase ASNA1 |
WRB | tryptophan-rich basic protein |
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Blenski, M.; Kehlenbach, R.H. Targeting of LRRC59 to the Endoplasmic Reticulum and the Inner Nuclear Membrane. Int. J. Mol. Sci. 2019, 20, 334. https://doi.org/10.3390/ijms20020334
Blenski M, Kehlenbach RH. Targeting of LRRC59 to the Endoplasmic Reticulum and the Inner Nuclear Membrane. International Journal of Molecular Sciences. 2019; 20(2):334. https://doi.org/10.3390/ijms20020334
Chicago/Turabian StyleBlenski, Marina, and Ralph H. Kehlenbach. 2019. "Targeting of LRRC59 to the Endoplasmic Reticulum and the Inner Nuclear Membrane" International Journal of Molecular Sciences 20, no. 2: 334. https://doi.org/10.3390/ijms20020334