Analysis of Kaposi's sarcoma-associated herpesvirus latent replication using a real-time polymerase chain reaction technique.
Kaposi's sarcoma-associated herpesvirus (KSHV) undergoes replication independently via latent and lytic pathways. Latent replication is mediated by latent-associated nuclear antigen (LANA), the sole viral trans element for genome maintenance and replication. According to previous studies, LANA tethers the KSHV genome to the host chromosome during latency and interacts with host factors to ensure proper latent replication. Studies using Southern blot experiments have revealed consistently that vector constructs containing the viral terminal repeat (TR) region as a cis element in latent replication are replicated in the presence of LANA. However, Southern blotting is a time-intensive, complicated technique that requires multiple reagents. In addition, it has a limited ability to detect slight changes in replication efficiency under different conditions owing to its relatively low sensitivity. In the current study, a real-time polymerase chain reaction method was developed for detecting transient KSHV replication and was found to be capable of further identifying several factors that affect latent replication. This technique should provide a useful tool for the detection of KSHV latent replication under various conditions, including overexpression of viral or cellular factors and chemical stimulation.
Department of Life Science, Dongguk University-Seoul, Seoul 100-715, South Korea.
Seho
S
Cha
Jun Hyeong
JH
Jang
Yejin
Y
Kim
Lee Rang
LR
Hwang
Taegun
T
Seo
D000956:Antigens, Viral / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000502:physiology*; D009687:Nuclear Proteins / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D014773:Virology / Q000379:methods*; D014779:Virus Replication
D000956:Antigens, Viral; D004279:DNA, Viral; D009687:Nuclear Proteins; C109540:latency-associated nuclear antigen
KSHV; Kaposi's sarcoma-associated herpesvirus; LANA; Latent replication; Real-time PCR; TR; latent-associated nuclear antigen; terminal repeat