Proteasome activity is essential for pollen tube emergence and growth; nevertheless, little is known about proteasome function at the molecular level. The objective of this study was to identify molecular targets and pathways which are directly/indirectly controlled by the proteasome during pollen germination. To this aim, changes in the proteome and phosphoproteome of Actinidia pollen, germinated in the presence of the proteasome inhibitor MG132, were investigated. Phosphoproteins were enriched by metal oxide/hydroxide affinity chromatography and phosphopeptides were further isolated using titanium ion (Ti4+) functional magnetic microparticles prior to liquid chromatography-tandem mass spectrometry analysis. Our results show that proteasome inhibition affects the phosphoproteome more profoundly than the proteome. Accordingly, the steady-state abundance of some kinases and phosphatases was changed and/or their phosphorylation status altered. Notably, affected proteins are involved in processes that are fundamental to pollen germination such as cytoskeletal organization, vesicular transport, cell wall synthesis and remodeling, protein synthesis, folding and degradation as well as energetic metabolism. Our data provide a molecular framework for the structural alterations observed when the proteasome is inhibited, contribute to the understanding of how proteasome activity regulates pollen germination, show the cross-talk between phosphorylation and proteasomal degradation and are a resource for further functional analyses.

Proteasome-mediated remodeling of the proteome and phosphoproteome during kiwifruit pollen germination

Scoccianti, Valeria;Ceccarini, Chiara;Crinelli, Rita
2019-01-01

Abstract

Proteasome activity is essential for pollen tube emergence and growth; nevertheless, little is known about proteasome function at the molecular level. The objective of this study was to identify molecular targets and pathways which are directly/indirectly controlled by the proteasome during pollen germination. To this aim, changes in the proteome and phosphoproteome of Actinidia pollen, germinated in the presence of the proteasome inhibitor MG132, were investigated. Phosphoproteins were enriched by metal oxide/hydroxide affinity chromatography and phosphopeptides were further isolated using titanium ion (Ti4+) functional magnetic microparticles prior to liquid chromatography-tandem mass spectrometry analysis. Our results show that proteasome inhibition affects the phosphoproteome more profoundly than the proteome. Accordingly, the steady-state abundance of some kinases and phosphatases was changed and/or their phosphorylation status altered. Notably, affected proteins are involved in processes that are fundamental to pollen germination such as cytoskeletal organization, vesicular transport, cell wall synthesis and remodeling, protein synthesis, folding and degradation as well as energetic metabolism. Our data provide a molecular framework for the structural alterations observed when the proteasome is inhibited, contribute to the understanding of how proteasome activity regulates pollen germination, show the cross-talk between phosphorylation and proteasomal degradation and are a resource for further functional analyses.
File in questo prodotto:
File Dimensione Formato  
JoP 2019.pdf

non disponibili

Tipologia: Versione editoriale
Licenza: Pubblico con Copyright
Dimensione 1.5 MB
Formato Adobe PDF
1.5 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Manuscript JoP 2018 accepted version.pdf

Open Access dal 11/02/2020

Descrizione: articolo principale
Tipologia: Versione referata/accettata
Licenza: Creative commons
Dimensione 640.87 kB
Formato Adobe PDF
640.87 kB Adobe PDF Visualizza/Apri
Figures S1-S4 JoP 2018.pdf

Open Access dal 11/02/2020

Descrizione: Figure supplementari
Tipologia: Versione referata/accettata
Licenza: Creative commons
Dimensione 580.7 kB
Formato Adobe PDF
580.7 kB Adobe PDF Visualizza/Apri
Supplementary table S1-S11.xlsx

Open Access dal 11/02/2020

Descrizione: Tabelle supplementari
Tipologia: Versione referata/accettata
Licenza: Creative commons
Dimensione 1.93 MB
Formato Microsoft Excel XML
1.93 MB Microsoft Excel XML Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11576/2663456
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 7
  • ???jsp.display-item.citation.isi??? 6
social impact