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SUMO Protocols pp 285-301 | Cite as

Inhibition of the SUMO Pathway by Gam1

  • Mariaelena Pozzebon
  • Chiara V. Segré
  • Susanna Chiocca
Protocol
Part of the METHODS IN MOLECULAR BIOLOGY™ book series (MIMB, volume 497)

Abstract

We have previously demonstrated that Gam1, an avian adenoviral protein inhibits sumoylation. By counteracting the SUMO pathway, Gam1 has a significant impact on virus-infected cells, but in isolation the inhibitory effects of the Gam1 protein can be exploited to intentionally manipulate the SUMO system in vivo or in vitro. Here we discuss in detail the techniques we use to inhibit the SUMO pathway using the Gam1 protein.

Keywords

Gam1 SAE1/SAE2 UBC9 SUMO SUMO E1 SUMO E2 inhibition sumoylation degradation 

Notes

Acknowledgments

We thank all IFOM-IEO campus facilities and all members of our laboratory, past and present. A special thank to Ario de Marco, Marisa Aliprandi and Annunziata Venuto for creating the inducible Gam1 expressing cell line, to Alfonso passafaro for generating the pMSCV-IRES-Gam1-GFP Plasmid and to Ronald T. Hay for kindly providing GST-SAE2/SAE1 and GST-SUMO-1 (1–97aa-C52A plasmids). This work was supported by grants from AIRC (Associazione Italiana per la Ricerca sul Cancro) to S.C., and from the Italian Ministry of Health.

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Mariaelena Pozzebon
    • 1
  • Chiara V. Segré
    • 1
  • Susanna Chiocca
    • 2
  1. 1.FIRC Institute of Molecular OncologyEuropean Institute of Oncology (IFOM-IEO)MilanItaly
  2. 2.Department of Experimental OncologyEuropean Institute of OncologyMilanItaly

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