title: Arabidopsis PCNAs form complexes with selected D-type cyclins.
abstract: Proliferating Cell Nuclear Antigen (PCNA) is a key nuclear protein of eukaryotic cells. It has been shown to form complexes with cyclin dependent kinases, cyclin dependent kinase inhibitors and the D-type cyclins which are involved in the cell cycle control. In Arabidopsis two genes coding for PCNA1 and PCNA2 proteins have been identified. In this study by analyzing Arabidopsis PCNA/CycD complexes we tested the possible functional differentiation of PCNA1/2 proteins in cell cycle control. Most out of the 10 cyclins investigated showed only nuclear localization except CycD2;1, CycD4;1, and CycD4;2 which were observed both in the nucleus and cytoplasm. Using the Y2H, BiFC and FLIM-FRET techniques we identified D-type cyclins which formed complexes with either PCNA1 or PCNA2. Among the candidates tested only CycD1;1, CycD3;1, and CycD3;3 were not detected in a complex with the PCNA proteins. Moreover, our results indicate that the formation of CycD3;2/PCNA and CycD4;1/PCNA complexes can be regulated by other as yet unidentified factor(s). Additionally, FLIM-FRET analyses suggested that in planta the distance between PCNA1/CycD4;1, PCNA1/CycD6;1, PCNA1/CycD7;1, and PCNA2/CycD4;2 proteins was shorter than that between PCNA2/CycD4;1, PCNA2/CycD6;1, PCNA2/CycD7;1, and PCNA1/CycD4;2 pairs. These data indicate that the nine amino acid differences between PCNA1 and PCNA2 have an impact on the architecture of Arabidopsis CycD/PCNA complexes.
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University Krakow, Poland ; The Bioremediation Department, Malopolska Centre of Biotechnology, Jagiellonian University Krakow, Poland.
authors.forename: Wojciech K
authors.initials: WK
authors.lastname: Strzalka
authors.affiliation: Department of Gene Expression, Faculty of Biology, Adam Mickiewicz University Poznan, Poland.
authors.forename: Chhavi
authors.initials: C
authors.lastname: Aggarwal
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University Krakow, Poland.
authors.forename: Weronika
authors.initials: W
authors.lastname: Krzeszowiec
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University Krakow, Poland.
authors.forename: Agata
authors.initials: A
authors.lastname: Jakubowska
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University Krakow, Poland.
authors.forename: Olga
authors.initials: O
authors.lastname: Sztatelman
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University Krakow, Poland.
authors.forename: Agnieszka K
authors.initials: AK
authors.lastname: Banas
keywords: Arabidopsis; D-type cyclins; DNA repair; DNA replication; PCNA; cell cycle