Function of deubiquitylating enzymes in the selective degradation of plasma membrane proteins in Arabidopsis thaliana

Vogel, Karin

Function of deubiquitylating enzymes in the selective degradation of plasma membrane proteins in Arabidopsis thaliana

Dateien

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Datum

2022

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Biologie

Publikationstyp

Dissertation

Publikationsstatus

Published

Zusammenfassung

Plants are sessile organisms that need to tightly regulate the abundance of plasma membrane (PM) proteins to react to different environmental stimuli. As ubiquitylation is the key signal for selective protein degradation, the stability of PM proteins is regulated by ubiquitylating as well as deubiquitylating enzymes (DUBs). In Arabidopsis thaliana, the metalloprotease DUBs AMSH1 and AMSH3 influence the stability of PM proteins. The exact nature of the proteins accumulating in the amsh1 and amsh3 mutants is, however, unknown. To expand the knowledge about AMSH function, potential AMSH1 targets were identified with an affinity purification and subsequent MS analysis. As we assumed that AMSH1 targets carry a K63-linked ubiquitin chain, proteins were purified with an ubiquitin-binding construct from amsh1 and wild-type seedlings. To identify potential AMSH1 targets, amsh1 and wild-type ubiquitylomes were compared. The observation that potential target proteins were enriched both in the wild-type and amsh1 background indicates that AMSH1 could stabilize some proteins whereas it destabilizes others. AMSH1 seems to be versatile DUB with different regulatory functions in the Arabidopsis cell. Selective protein degradation pathways consist of various steps that need to be tightly regulated by ubiquitylation and deubiquitylation. So it is likely that there are multiple Arabidopsis DUBs which can influence the stability of PM proteins. The DUBs OTU11 and OTU12 are localized at the PM in Arabidopsis root cells and could therefore influence the selective degradation of PM proteins. OTU11 and OTU12 bind directly to PM-localized phosphatidylinositol phosphates (PIPs) in vitro. As the weak in vitro activity of OTU11 against K63-linked ubiquitin was enhanced by binding to liposomes, the catalytic activity of OTU11 seems to be tightly connected to its membrane localization. Furthermore, OTU11 and OTU12 had an influence on the root length of Arabidopsis seedlings, stabilized the artificial endocytosis substrate PMA-GFP-UB at the PM, and had an effect on the endocytic degradation of the PM protein PIN2-GFP. Altogether, the experimental data shows that OTU11 and OTU12 are PM localized DUBs which could have fine-tuning effect on the abundance of PM proteins in Arabidopsis and identified a novel DUB function in the selective protein degradation pathways of PM proteins in Arabidopsis.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Zitieren

ISO 690

VOGEL, Karin, 2022. Function of deubiquitylating enzymes in the selective degradation of plasma membrane proteins in Arabidopsis thaliana [Dissertation]. Konstanz: University of Konstanz

BibTex

@phdthesis{Vogel2022Funct-59584,
  year={2022},
  title={Function of deubiquitylating enzymes in the selective degradation of plasma membrane proteins in Arabidopsis thaliana},
  author={Vogel, Karin},
  address={Konstanz},
  school={Universität Konstanz}
}

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Prüfungsdatum der Dissertation

June 3, 2022

Hochschulschriftenvermerk

Konstanz, Univ., Diss., 2022

Universitätsbibliographie

Ja