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  1. Article: Inhibition of the SUMO pathway by Gam1.

    Pozzebon, Mariaelena / Segré, Chiara V / Chiocca, Susanna

    Methods in molecular biology (Clifton, N.J.)

    2009  Volume 497, Page(s) 285–301

    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 ... ...

    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.
    MeSH term(s) Animals ; Cells, Cultured ; Clinical Laboratory Techniques ; Humans ; Protein Processing, Post-Translational/drug effects ; Recombinant Proteins/isolation & purification ; Recombinant Proteins/pharmacology ; SUMO-1 Protein/antagonists & inhibitors ; SUMO-1 Protein/metabolism ; Signal Transduction/drug effects ; Viral Proteins/isolation & purification ; Viral Proteins/pharmacology
    Chemical Substances CELO protein, adenovirus ; Recombinant Proteins ; SUMO-1 Protein ; Viral Proteins
    Language English
    Publishing date 2009
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ISSN 1064-3745
    ISSN 1064-3745
    DOI 10.1007/978-1-59745-566-4_19
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Inhibition of S-phase progression triggered by UVA-induced ROS does not require a functional DNA damage checkpoint response in mammalian cells.

    Girard, Pierre-Marie / Pozzebon, Mariaelena / Delacôte, Fabien / Douki, Thierry / Smirnova, Violetta / Sage, Evelyne

    DNA repair

    2008  Volume 7, Issue 9, Page(s) 1500–1516

    Abstract: Ultraviolet A (UVA) radiation represents more than 90% of the UV spectrum reaching Earth's surface. Exposure to UV light, especially the UVA part, induces the formation of photoexcited states of cellular photosensitizers with subsequent generation of ... ...

    Abstract Ultraviolet A (UVA) radiation represents more than 90% of the UV spectrum reaching Earth's surface. Exposure to UV light, especially the UVA part, induces the formation of photoexcited states of cellular photosensitizers with subsequent generation of reactive oxygen species (ROS) leading to damages to membrane lipids, proteins and nucleic acids. Although UVA, unlike UVC and UVB, is poorly absorbed by DNA, it inhibits cell cycle progression, especially during S-phase. In the present study, we examined the role of the DNA damage checkpoint response in UVA-induced inhibition of DNA replication. We provide evidence that UVA delays S-phase in a dose dependent manner and that UVA-irradiated S-phase cells accumulate in G2/M. We show that upon UVA irradiation ATM-, ATR- and p38-dependent signalling pathways are activated, and that Chk1 phosphorylation is ATR/Hus1 dependent while Chk2 phosphorylation is ATM dependent. To assess for a role of these pathways in UVA-induced inhibition of DNA replication, we investigated (i) cell cycle progression of BrdU labelled S-phase cells by flow cytometry and (ii) incorporation of [methyl-(3)H]thymidine, as a marker of DNA replication, in ATM, ATR and p38 proficient and deficient cells. We demonstrate that none of these pathways is required to delay DNA replication in response to UVA, thus ruling out a role of the canonical S-phase checkpoint response in this process. On the contrary, scavenging of UVA-induced reactive oxygen species (ROS) by the antioxidant N-acetyl-L-cystein or depletion of vitamins during UVA exposure significantly restores DNA synthesis. We propose that inhibition of DNA replication is due to impaired replication fork progression, rather as a consequence of UVA-induced oxidative damage to protein than to DNA.
    MeSH term(s) Ataxia Telangiectasia Mutated Proteins ; Cell Cycle/drug effects ; Cell Cycle/radiation effects ; Cell Cycle Proteins/metabolism ; Cell Line, Transformed ; DNA/biosynthesis ; DNA Damage ; DNA Replication/drug effects ; DNA Replication/radiation effects ; DNA-Binding Proteins/metabolism ; Humans ; Phosphatidylinositol 3-Kinases/metabolism ; Protein-Serine-Threonine Kinases/metabolism ; Reactive Oxygen Species/pharmacology ; S Phase/drug effects ; S Phase/radiation effects ; Signal Transduction ; Tumor Suppressor Proteins/metabolism ; Ultraviolet Rays ; p38 Mitogen-Activated Protein Kinases/metabolism
    Chemical Substances Cell Cycle Proteins ; DNA-Binding Proteins ; Reactive Oxygen Species ; Tumor Suppressor Proteins ; DNA (9007-49-2) ; Phosphatidylinositol 3-Kinases (EC 2.7.1.-) ; ATM protein, human (EC 2.7.11.1) ; ATR protein, human (EC 2.7.11.1) ; Ataxia Telangiectasia Mutated Proteins (EC 2.7.11.1) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; p38 Mitogen-Activated Protein Kinases (EC 2.7.11.24)
    Language English
    Publishing date 2008-09-01
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2071608-4
    ISSN 1568-7856 ; 1568-7864
    ISSN (online) 1568-7856
    ISSN 1568-7864
    DOI 10.1016/j.dnarep.2008.05.004
    Database MEDical Literature Analysis and Retrieval System OnLINE

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