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Published on: May 3, 2018
Protein Kinase D2 Regulates GRASP65 Phosphorylation and Golgi Ribbon Unlinking During G2/M Transition
Inmaculada Ayala1, Daniela Spano1, Antonino Colanzi1
1Institute of Endotypes in Oncology, Metabolism and Immunology "G. Salvatore" (IEOMI), National Research Council (CNR), Via P. Castellino 111, 80131 Naples, Italy.
Protein kinase D2 (PKD2) regulates Golgi ribbon unlinking by phosphorylating GRASP65, a key step for cell division. This discovery clarifies Golgi disassembly during the G2/M transition.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Golgi complex remodeling is crucial for cell cycle progression.
- GRASP65 phosphorylation by JNK2 is essential for Golgi ribbon unlinking.
- Upstream regulation of GRASP65 phosphorylation remained unclear.
Purpose of the Study:
- Identify upstream regulators of GRASP65 phosphorylation.
- Investigate the role of PKD2 in Golgi unlinking.
- Develop tools to study Golgi disassembly.
Main Methods:
- Generated and validated a phospho-specific antibody for human GRASP65 (pS274).
- Utilized PKD2 inhibition and depletion studies.
- Employed PKD2-activating stimuli (phorbol esters, nocodazole).
Main Results:
- PKD2 is identified as a critical upstream regulator of GRASP65 phosphorylation at S274.
- PKD2 activity directly correlates with GRASP65 phosphorylation and Golgi unlinking.
- PKD2 inhibition or depletion leads to decreased Golgi unlinking and delayed G2/M transition.
- PKD2-activating stimuli enhance GRASP65 phosphorylation in a PKD2-dependent manner.
Conclusions:
- PKD2 is a key regulator of the JNK2-GRASP65 signaling axis controlling Golgi disassembly.
- PKD2-mediated GRASP65 phosphorylation is essential for Golgi unlinking at G2/M transition.
- The phospho-specific GRASP65 antibody is a valuable tool for studying upstream signaling in Golgi unlinking.
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