Characterization of human herpesvirus 6 variant B immediate-early 1 protein modifications by small ubiquitin-related modifiers.
The human herpesvirus 6 (HHV-6) immediate-early (IE) 1 protein undergoes SUMOylation events during the infectious process. In the present work, we report that Lys-802 (K-802) of IE1 from HHV-6 variant B is the only target residue capable of conjugation to SUMO-1/SMT3C/Sentrin-1, SUMO-2/SMT3A/Sentrin-3 or SUMO-3/SMT3B/Sentrin-2 as determined by transfection and in vitro SUMOylation experiments. PolySUMOylated forms of IE1 were also observed, suggesting that SUMO branching occurs at the K-802 residue. Overexpression of SUMO-1, -2 and -3 led to an overall increase in IE1 levels, irrespective of K-802. The SUMO residues could be efficiently removed by incubating SUMOylated IE1 with SENP1, a recently identified SUMO peptidase. SUMOylation-deficient mutants of IE1 co-localized with nuclear promyelocytic leukaemia protein (PML) oncogenic domains (PODs) as efficiently as WT IE1, indicating that POD targeting is independent of IE1 SUMOylation status. However, in contrast to infection, PODs did not aggregate in IE1B-transfected cells, suggesting that other viral proteins are involved in the process. Transactivation studies indicated that IE1, in combination with IE2, could efficiently transactivate diverse promoters, independent of its SUMOylation status. Overall, the results presented provide a detailed biochemical characterization of post-translational modifications of the HHV-6 IE1 protein by SUMO peptides, contributing to our understanding of the complex interactions between herpesviruses and the SUMO-conjugation pathway.
Gravel
Annie
A
Laboratory of Virology, Rheumatology and Immunology Research Center, Room T1-49, CHUL Research Center and Faculty of Medicine, Laval University, 2705 Laurier Blvd, Sainte-Foy, Quebec, Canada G1V 4G2.
Dion
Valérie
V
Laboratory of Virology, Rheumatology and Immunology Research Center, Room T1-49, CHUL Research Center and Faculty of Medicine, Laval University, 2705 Laurier Blvd, Sainte-Foy, Quebec, Canada G1V 4G2.
Cloutier
Nathalie
N
Laboratory of Virology, Rheumatology and Immunology Research Center, Room T1-49, CHUL Research Center and Faculty of Medicine, Laval University, 2705 Laurier Blvd, Sainte-Foy, Quebec, Canada G1V 4G2.
Gosselin
Jean
J
Laboratory of Viral Immunology, Rheumatology and Immunology Research Center, Room T1-49, CHUL Research Center and Faculty of Medicine, Laval University, 2705 Laurier Blvd, Sainte-Foy, Quebec, Canada G1V 4G2.
Flamand
Louis
L
Laboratory of Virology, Rheumatology and Immunology Research Center, Room T1-49, CHUL Research Center and Faculty of Medicine, Laval University, 2705 Laurier Blvd, Sainte-Foy, Quebec, Canada G1V 4G2.
D000818:Animals; D002460:Cell Line; D015654:Herpesvirus 6, Human / Q000378:metabolism*; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D008239:Lysine; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*
D017874:Immediate-Early Proteins; D010750:Phosphoproteins; D025841:Small Ubiquitin-Related Modifier Proteins; C458509:immediate-early 1 protein, human herpesvirus 6; D008239:Lysine