RESUMEN
Since the identification of the antisense protein of HTLV-2 (APH-2) and the demonstration that APH-2 mRNA is expressed in vivo in most HTLV-2 carriers, much effort has been dedicated to the elucidation of similarities and/or differences between APH-2 and HBZ, the antisense protein of HTLV-1. Similar to HBZ, APH-2 negatively regulates HTLV-2 transcription. However, it does not promote cell proliferation. In contrast to HBZ, APH-2 half-life is very short. Here, we show that APH-2 is addressed to PML nuclear bodies in T-cells, as well as in different cell types. Covalent SUMOylation of APH-2 is readily detected, indicating that APH-2 might be addressed to the PML nuclear bodies in a SUMO-dependent manner. We further show that silencing of PML increases expression of APH-2, while expression of HBZ is unaffected. On the other hand, SUMO-1 overexpression leads to a specific loss of APH-2 expression that is restored upon proteasome inhibition. Furthermore, the carboxy-terminal LAGLL motif of APH-2 is responsible for both the targeting of the protein to PML nuclear bodies and its short half-life. Taken together, these observations indicate that natural APH-2 targeting to PML nuclear bodies induces proteasomal degradation of the viral protein in a SUMO-dependent manner. Hence, this study deciphers the molecular and cellular bases of APH-2 short half-life in comparison to HBZ and highlights key differences in the post-translational mechanisms that control the expression of both proteins.
Asunto(s)
Virus Linfotrópico T Tipo 2 Humano/metabolismo , Cuerpos de Inclusión Intranucleares/metabolismo , Proteína de la Leucemia Promielocítica/metabolismo , Proteínas Virales/química , Proteínas Virales/metabolismo , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Regulación Viral de la Expresión Génica , Semivida , Células HeLa , Virus Linfotrópico T Tipo 2 Humano/genética , Humanos , Células Jurkat , Complejo de la Endopetidasa Proteasomal/metabolismo , Proteolisis , Proteínas de los Retroviridae/metabolismo , Proteína SUMO-1/metabolismo , Sumoilación , Transcripción GenéticaRESUMEN
We recently discovered the antisense protein of human T-cell leukemia virus (HTLV) type 2 (APH-2), whose messenger RNA is encoded by the antisense strand of the HTLV-2 genome. We quantified proviral load, level of tax, and APH-2 in a series of blood samples obtained from a cohort of HTLV-2 carriers. We determined whether APH-2 promotes cell proliferation. APH-2 was detectable in most samples tested and was correlated with proviral load. APH-2 levels were not correlated with lymphocyte count in vivo, consistent with the inability of APH-2 to promote cell proliferation in vitro. APH-2 does not promote cell proliferation and does not cause lymphocytosis.
Asunto(s)
Infecciones por HTLV-II/virología , Virus Linfotrópico T Tipo 2 Humano/metabolismo , Linfocitosis/virología , Provirus/metabolismo , Proteínas de los Retroviridae/sangre , Carga Viral , Adulto , Anciano , Anciano de 80 o más Años , Infecciones Asintomáticas , Estudios de Cohortes , Femenino , Productos del Gen tax/sangre , Infecciones por HTLV-II/sangre , Infecciones por HTLV-II/complicaciones , Humanos , Modelos Lineales , Masculino , Persona de Mediana EdadRESUMEN
Human T-cell leukemia virus type 1 (HTLV-1) and type 2 (HTLV-2) retroviruses infect T lymphocytes. The minus strand of the HTLV-1 genome encodes HBZ, a protein that could play a role in the development of leukemia in infected patients. Herein, we demonstrate that the complementary strand of the HTLV-2 genome also encodes a protein that we named APH-2 for "antisense protein of HTLV-2." APH-2 mRNA is spliced, polyadenylated, and initiates in the 3'-long terminal repeat at different positions. This transcript was detected in all HTLV-2-infected cell lines and short-term culture of lymphocytes obtained from HTLV-2 African patients tested and in 4 of 15 HTLV-2-infected blood donors. The APH-2 protein is 183 amino acids long, is localized in the cell nucleus, and is detected in vivo. Despite the lack of a consensus basic leucine zipper domain, APH-2 interacts with cyclic adenosine monophosphate-response element binding protein (CREB) and represses Tax2-mediated transcription in Tax2-expressing cells and in cells transfected with an HTLV-2 molecular clone. Altogether, our results demonstrate the existence of an antisense strand-encoded protein in HTLV-2, which could represent an important player in the development of disorders, such as lymphocytosis, which is frequently observed in HTLV-2 patients.
Asunto(s)
Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Productos del Gen tax/genética , Virus Linfotrópico T Tipo 2 Humano/fisiología , Empalme del ARN/genética , ARN sin Sentido/genética , Transcripción Genética , Proteínas Virales/genética , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Bases , Northern Blotting , Western Blotting , Femenino , Técnica del Anticuerpo Fluorescente , Regulación Viral de la Expresión Génica , Productos del Gen tax/metabolismo , Humanos , Técnicas para Inmunoenzimas , Inmunoprecipitación , Células Jurkat , Luciferasas/genética , Luciferasas/metabolismo , Persona de Mediana Edad , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , ARN sin Sentido/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Ácido Nucleico , Transfección , Proteínas Virales/metabolismoRESUMEN
Despite being discovered in animals in the early 20th century, the scientific interest in retroviruses was boosted with the discovery of human retroviruses (human T-leukemia/lymphoma virus [HTLV] and HIV), which are responsible for significant morbidity and mortality. HTLV was identified more than 25 years ago as the etiological agent of adult T-cell leukemia/lymphoma. It was then shown to be a complex retrovirus, given that it not only encodes the characteristic retroviral Gag, Pol and Env proteins, but also regulatory and accessory proteins. Since the first studies documenting the role of these proteins in viral expression, the picture has become increasingly more complex. Indeed, owing to the limited size of its genome that contains overlapping open-reading frames, HTLV has evolved unique ways to regulate its expression. Retroviral expression was originally thought to be mainly controlled through the regulation of transcription from the 5 long-terminal repeats, but we now know that the 3 long-terminal repeats also serve as promoters. Regulation of splicing and mRNA export, and post-translational modifications of viral protein also play a major role. This review discusses the latest insights gained into the field of HTLV gene expression.