Covalent modification by SUMO is required for efficient disruption of PML oncogenic domains by Kaposi's sarcoma-associated herpesvirus latent protein LANA2.
The multifunctional Kaposi's sarcoma-associated herpesvirus (KSHV) latent protein latency-associated nuclear antigen 2 (LANA2) has a critical role in KSHV-induced B-cell malignancies. LANA2 increases the level of small ubiquitin-like modifier (SUMO)2-ubiquitin-modified PML and induces the disruption of PML oncogenic domains (PODs) by a process that requires a non-covalent SUMO interaction domain (SIM) in LANA2. We now demonstrate that LANA2 is covalently conjugated to SUMO1 and SUMO2 both in vitro and in latently KSHV-infected B-cells. We show that a LANA2 SIM mutant exhibits a slightly altered sumoylation pattern, which suggests that non-covalent SUMO interactions represent a mechanism for determining SUMO substrate recognition and modification. In addition, several lysine residues were mapped as SUMO conjugation sites. A sumoylation-deficient mutant shows impaired ability to induce disruption of PODs, which suggests that either directly bound or covalently conjugated SUMO moieties may act as a bridge for interaction between LANA2 and other SUMO-modified or SUMO-interacting proteins required for disruption of PODs.
Marcos-Villar
Laura
L
Dept Microbiología II, Fac. Farmacia, Universidad Complutense de Madrid, Pza Ramón y Cajal, 28040 Madrid, Spain.
Campagna
Michela
M
Lopitz-Otsoa
Fernando
F
Gallego
Pedro
P
González-Santamaría
José
J
González
Dolores
D
Rodriguez
Manuel S
MS
Rivas
Carmen
C
D000956:Antigens, Viral / Q000378:metabolism*; D001402:B-Lymphocytes / Q000821:virology; D002460:Cell Line; D019288:Herpesvirus 8, Human / Q000472:pathogenicity*; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D025842:SUMO-1 Protein / Q000378:metabolism*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D058207:Sumoylation; D017735:Virus Latency
D000956:Antigens, Viral; D009687:Nuclear Proteins; D025842:SUMO-1 Protein; C117909:SUMO2 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; C109540:latency-associated nuclear antigen