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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
Published on: May 3, 2018
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Arabidopsis PCNAs form complexes with selected D-type cyclins.
Wojciech K Strzalka1, Chhavi Aggarwal2, Weronika Krzeszowiec3
1Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University Krakow, Poland ; The Bioremediation Department, Malopolska Centre of Biotechnology, Jagiellonian University Krakow, Poland.
Frontiers in Plant Science
|September 18, 2015
Summary
Arabidopsis Proliferating Cell Nuclear Antigen (PCNA) proteins PCNA1 and PCNA2 show functional differentiation in cell cycle control. Nine amino acid differences impact the architecture of PCNA/CycD complexes, influencing plant cell proliferation.
Area of Science:
- Plant Molecular Biology
- Cell Cycle Regulation
- Protein Interactions
Background:
- Proliferating Cell Nuclear Antigen (PCNA) is crucial for eukaryotic cell cycle control.
- In Arabidopsis, two PCNA genes (PCNA1 and PCNA2) have been identified.
- PCNA proteins interact with cyclins and cyclin-dependent kinases involved in cell cycle regulation.
Purpose of the Study:
- To investigate the functional differentiation of Arabidopsis PCNA1 and PCNA2 proteins.
- To analyze the formation and architecture of PCNA/CycD complexes in Arabidopsis.
- To understand the role of PCNA isoforms in plant cell cycle control.
Main Methods:
- Yeast Two-Hybrid (Y2H) assays
- Bimolecular Fluorescence Complementation (BiFC)
- Fluorescence Lifetime Imaging Microscopy-Förster Resonance Energy Transfer (FLIM-FRET)
- Analysis of Arabidopsis D-type cyclins and PCNA interactions
Main Results:
- Identified specific D-type cyclins that form complexes with either PCNA1 or PCNA2.
- Found that CycD1;1, CycD3;1, and CycD3;3 do not form complexes with PCNA proteins.
- Observed that CycD3;2/PCNA and CycD4;1/PCNA complex formation may be regulated by other factors.
- FLIM-FRET data indicated distinct distances between PCNA/CycD pairs, suggesting structural differences.
- Nine amino acid variations between PCNA1 and PCNA2 influence the architecture of Arabidopsis CycD/PCNA complexes.
Conclusions:
- Arabidopsis PCNA1 and PCNA2 proteins exhibit functional differentiation in cell cycle control.
- The specific interactions and complex architecture are influenced by variations between PCNA isoforms.
- These findings contribute to understanding the nuanced regulation of the plant cell cycle.
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