TPX2 (Targeting Protein for Xklp2) is an evolutionary conserved microtubule-associated protein important for microtubule nucleation and mitotic spindle assembly. The protein was described as an activator of the mitotic kinase Aurora A in humans and the Arabidopsis AURORA1 (AUR1) kinase. In contrast to animal genomes that encode only one TPX2 gene, higher plant genomes encode a family with several TPX2-LIKE gene members (TPXL). TPXL genes of Arabidopsis can be divided into two groups. Group A proteins (TPXL2, 3, 4, and 8) contain Aurora binding and TPX2_importin domains, while group B proteins (TPXL1, 5, 6, and 7) harbor an Xklp2 domain. Canonical TPX2 contains all the above-mentioned domains. We confirmed using in vitro kinase assays that the group A proteins contain a functional Aurora kinase binding domain. Transient expression of Arabidopsis TPX2-like proteins in Nicotiana benthamiana revealed preferential localization to microtubules and nuclei. Co-expression of AUR1 together with TPX2-like proteins changed the localization of AUR1, indicating that these proteins serve as targeting factors for Aurora kinases. Taken together, we visualize the various localizations of the TPX2-LIKE family in Arabidopsis as a proxy to their functional divergence and provide evidence of their role in the targeted regulation of AUR1 kinase activity.
- MeSH
- Arabidopsis cytologie genetika metabolismus MeSH
- kinasy aurora metabolismus MeSH
- mikrotubuly metabolismus MeSH
- proteinové domény MeSH
- proteiny asociované s mikrotubuly analýza genetika metabolismus MeSH
- proteiny huseníčku analýza genetika metabolismus MeSH
- rostlinné geny MeSH
- sekvence aminokyselin MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
Fluorescence proteins changing spectral properties after exposure to light with a specific wavelength have recently become outstanding aids in the study of intracellular protein dynamics. Herein we show using Arabidopsis SYNAPTOTAGMIN 1 as a model protein that the Dendra2 green to red photoconvertible protein tag in combination with confocal scanning laser microscopy is a useful tool to study membrane protein intracellular dynamics.
- MeSH
- Arabidopsis chemie cytologie ultrastruktura MeSH
- fluorescenční barviva analýza MeSH
- intracelulární membrány chemie ultrastruktura MeSH
- konfokální mikroskopie metody MeSH
- kořeny rostlin chemie ultrastruktura MeSH
- luminescentní proteiny analýza MeSH
- proteiny huseníčku analýza MeSH
- světlo MeSH
- synaptotagmin I analýza MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Actin cytoskeleton is a vital cellular structure primarily known for controlling cell integrity, division and expansion. Here we present a proteomic dissection of Arabidopsis roots treated by actin depolymerizing agent latrunculin B. Pharmacological disintegration of the actin cytoskeleton by latrunculin B caused downregulation of several proteins involved in the actin organization and dynamics. Moreover, this approach helped to identify new protein candidates involved in gene transcription, due to the altered abundance of proteins involved in mRNA nuclear export. Finally, latrunculin B negatively affected the abundance of abscisic acid (ABA) responsive proteins. SIGNIFICANCE: This article substantially contributes to the current knowledge about the importance of actin organization and dynamics in proteome remodelling. We employed gel based and gel free proteomic analyses and identified several new protein candidates and protein networks linking actin dynamics to the gene transcription and to the ABA response in Arabidopsis.
- MeSH
- aktiny chemie metabolismus MeSH
- Arabidopsis chemie MeSH
- bicyklické sloučeniny heterocyklické farmakologie MeSH
- kořeny rostlin chemie MeSH
- kyselina abscisová fyziologie MeSH
- mikrofilamenta účinky léků MeSH
- polymerizace účinky léků MeSH
- proteiny huseníčku analýza chemie metabolismus MeSH
- proteom analýza účinky léků MeSH
- proteomika metody MeSH
- thiazolidiny farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
