TY - JOUR
T1 - SUMOylation of SAMHD1 at Lysine 595 is required for HIV-1 restriction in non-cycling cells
AU - Martinat, Charlotte
AU - Cormier, Arthur
AU - Tobaly-Tapiero, Joëlle
AU - Palmic, Noé
AU - Casartelli, Nicoletta
AU - Mahboubi, Bijan
AU - Coggins, Si’Ana A.A.
AU - Buchrieser, Julian
AU - Persaud, Mirjana
AU - Diaz-Griffero, Felipe
AU - Espert, Lucile
AU - Bossis, Guillaume
AU - Lesage, Pascale
AU - Schwartz, Olivier
AU - Kim, Baek
AU - Margottin-Goguet, Florence
AU - Saïb, Ali
AU - Zamborlini, Alessia
N1 - Funding Information:
The authors thank H. de Thé and V.Lallemand-Breitenbach for discussion, A. Amara, and X. Carnec for critical reading of the manuscript. The authors thank M. Benkirane (IGH, Montpellier, France), N. Manel (I. Curie, Paris, France), A. Puissant (INSERM U944) and R. Hay (University of Dundee, Dundee, UK) for reagents. We are grateful to the Core facility of IRSL, Yasmine Khalil, and Renaud Batrin for technical support. The Wellcome Trust WTISSF121302 and the Oxford Martin School LC0910-004 (James Martin Stem Cell Facility Oxford). The iPS cell line used in this study was originally generated from a donor sample supplied by the Oxford Parkinson’s Disease Center (OPDC) study (funded by the Monument Trust Discovery Award from Parkinson’s UK, a charity registered in England and Wales (2581970) and in Scotland (SC037554), with the support of the National Institute for Health Research (NIHR) Oxford Biomedical Research Center based at Oxford University Hospitals NHS Trust and University of Oxford, and the NIHR Comprehensive Local Research Network), and was reprogrammed within StemBANCC (supported by the Innovative Medicines Initiative Joint Undertaking under grant agreement number 115439, resources of which are composed of financial contribution from the European Union’s Seventh Framework Program (FP7/ 2007e2013) and EFPIA companies’ in kind contribution). This work was supported by Sidaction (grant 2018-1-AEQ-12075 to A.Z.), Sidaction/FRM (grant VIH2016126003 to A.Z.), EU FP7 [HEALTH-2012-INNOVATION-1 ‘HIVINNOV’] (Grant no. 305137 to A.S. and A.Z.). C.M. was supported by fellowships from the French “Ministère de la Recherche et de l’Innovation” and Sidaction. Some experiments were performed in the laboratory of B.K., supported by NIH (grant AI136581 and AI150451 to B.K.).
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12/1
Y1 - 2021/12/1
N2 - SAMHD1 is a cellular triphosphohydrolase (dNTPase) proposed to inhibit HIV-1 reverse transcription in non-cycling immune cells by limiting the supply of the dNTP substrates. Yet, phosphorylation of T592 downregulates SAMHD1 antiviral activity, but not its dNTPase function, implying that additional mechanisms contribute to viral restriction. Here, we show that SAMHD1 is SUMOylated on residue K595, a modification that relies on the presence of a proximal SUMO-interacting motif (SIM). Loss of K595 SUMOylation suppresses the restriction activity of SAMHD1, even in the context of the constitutively active phospho-ablative T592A mutant but has no impact on dNTP depletion. Conversely, the artificial fusion of SUMO2 to a non-SUMOylatable inactive SAMHD1 variant restores its antiviral function, a phenotype that is reversed by the phosphomimetic T592E mutation. Collectively, our observations clearly establish that lack of T592 phosphorylation cannot fully account for the restriction activity of SAMHD1. We find that SUMOylation of K595 is required to stimulate a dNTPase-independent antiviral activity in non-cycling immune cells, an effect that is antagonized by cyclin/CDK-dependent phosphorylation of T592 in cycling cells.
AB - SAMHD1 is a cellular triphosphohydrolase (dNTPase) proposed to inhibit HIV-1 reverse transcription in non-cycling immune cells by limiting the supply of the dNTP substrates. Yet, phosphorylation of T592 downregulates SAMHD1 antiviral activity, but not its dNTPase function, implying that additional mechanisms contribute to viral restriction. Here, we show that SAMHD1 is SUMOylated on residue K595, a modification that relies on the presence of a proximal SUMO-interacting motif (SIM). Loss of K595 SUMOylation suppresses the restriction activity of SAMHD1, even in the context of the constitutively active phospho-ablative T592A mutant but has no impact on dNTP depletion. Conversely, the artificial fusion of SUMO2 to a non-SUMOylatable inactive SAMHD1 variant restores its antiviral function, a phenotype that is reversed by the phosphomimetic T592E mutation. Collectively, our observations clearly establish that lack of T592 phosphorylation cannot fully account for the restriction activity of SAMHD1. We find that SUMOylation of K595 is required to stimulate a dNTPase-independent antiviral activity in non-cycling immune cells, an effect that is antagonized by cyclin/CDK-dependent phosphorylation of T592 in cycling cells.
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UR - http://www.scopus.com/inward/citedby.url?scp=85111577085&partnerID=8YFLogxK
U2 - 10.1038/s41467-021-24802-5
DO - 10.1038/s41467-021-24802-5
M3 - Article
C2 - 34321470
AN - SCOPUS:85111577085
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 4582
ER -
