The latency-associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus transactivates the telomerase reverse transcriptase promoter.
Telomerase is a multi-subunit ribonucleoprotein holoenzyme that stabilizes telomere length through the addition of new repeat sequence to the ends of chromosomes. Telomerase reverse transcriptase is the subunit of this complex responsible for the enzymatic activity of telomerase. Expression of the reverse transcriptase is regulated at the level of transcription through the action of transcription factors that target its promoter. Most Kaposi's sarcoma tumor cells are latently infected with the Kaposi's sarcoma-associated herpesvirus, and the constitutive expression of a viral-encoded latency-associated nuclear antigen has been shown to be important for the maintenance of the viral episome. The proliferative nature of Kaposi's sarcoma suggests that this antigen may also play a critical role in viral-mediated oncogenesis. In this study telomerase reverse transcriptase promoter elements cloned into a luciferase reporter plasmid were analyzed to determine the ability of the latency-associated nuclear antigen to regulate transcription. The latency-associated nuclear antigen transactivated the full-length promoter in 293T, 293, and BJAB cell lines. Furthermore, truncation promoter studies implicated sequence from -130 to +5 in viral-mediated activation. This region contains five Sp1 transcription factor-binding sites. Electrophoretic mobility shift assays indicated that the latency-associated nuclear antigen targets and affects the Sp1-DNA complex in the context of BJAB nuclear extracts.
Knight
J S
JS
Medical Scientist Training Program, University of Michigan Medical Center CCGC 3217, Ann Arbor, Michigan 48109-0934, USA.
Cotter
M A
MA
Robertson
E S
ES
D000956:Antigens, Viral; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000276:immunology; D002472:Cell Transformation, Viral / Q000276:immunology; D004273:DNA, Neoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins; D015654:Herpesvirus 6, Human / Q000276:immunology*; D006801:Humans; D009687:Nuclear Proteins / Q000502:physiology*; D011401:Promoter Regions, Genetic; D012313:RNA; D012514:Sarcoma, Kaposi / Q000821:virology*; D016329:Sp1 Transcription Factor / Q000378:metabolism; D019098:Telomerase / Q000235:genetics*; D015533:Transcriptional Activation / Q000502:physiology*; D014407:Tumor Cells, Cultured
D000956:Antigens, Viral; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016329:Sp1 Transcription Factor; C109540:latency-associated nuclear antigen; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase