title: New insights on the transcriptional regulation of CD69 gene through a potent enhancer located in the conserved non-coding sequence 2.
abstract: The CD69 type II C-type lectin is one of the earliest indicators of leukocyte activation acting in lymphocyte migration and cytokine secretion. CD69 expression in hematopoietic lineage undergoes rapid changes depending on the cell-lineage, the activation state or the localization of the cell where it is expressed, suggesting a complex and tightly controlled regulation. Here we provide new insights on the transcriptional regulation of CD69 gene in mammal species. Through in silico studies, we analyzed several regulatory features of the 4 upstream conserved non-coding sequences (CNS 1-4) previously described, confirming a major function of CNS2 in the transcriptional regulation of CD69. In addition, multiple transcription binding sites are identified in the CNS2 region by DNA cross-species conservation analysis. By functional approaches we defined a core region of 226bp located within CNS2 as the main enhancer element of CD69 transcription in the hematopoietic cells analyzed. By chromatin immunoprecipitation, binding of RUNX1 to the core-CNS2 was shown in a T cell line. In addition, we found an activating but not essential role of RUNX1 in CD69 gene transcription by site-directed mutagenesis and RNA silencing, probably through the interaction with this potent enhancer specifically in the hematopoietic lineage. In summary, in this study we contribute with new evidences to the landscape of the transcriptional regulation of the CD69 gene.
authors: Activación Immunológica Group, Carlos III National Health Institute (ISCIII), Majadahonda, Spain.
authors: Teresa
authors: T
authors: Laguna
authors: Activación Immunológica Group, Carlos III National Health Institute (ISCIII), Majadahonda, Spain.
authors: Laura
authors: L
authors: Notario
authors: Laboratory of Genetics, Division of Oncology, CIMA, University of Navarra, Pamplona, Spain.
authors: Raffaella
authors: R
authors: Pippa
authors: Activación Immunológica Group, Carlos III National Health Institute (ISCIII), Majadahonda, Spain.
authors: Miguel G
authors: MG
authors: Fontela
authors: Activación Immunológica Group, Carlos III National Health Institute (ISCIII), Majadahonda, Spain.
authors: Berta N
authors: BN
authors: Vázquez
authors: Laboratory of Genetics, Division of Oncology, CIMA, University of Navarra, Pamplona, Spain.
authors: Miren
authors: M
authors: Maicas
authors: Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain.
authors: Noemí
authors: N
authors: Aguilera-Montilla
authors: Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain.
authors: Ángel L
authors: ÁL
authors: Corbí
authors: Laboratory of Genetics, Division of Oncology, CIMA, University of Navarra, Pamplona, Spain.
authors: María D
authors: MD
authors: Odero
authors: Activación Immunológica Group, Carlos III National Health Institute (ISCIII), Majadahonda, Spain. Electronic address: lauzurica@isciii.es.
authors: Pilar
authors: P
authors: Lauzurica
mesh_terms: D024506:5' Flanking Region; D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D050676:Core Binding Factor Alpha 2 Subunit / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D037181:Lectins, C-Type / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D014158:Transcription, Genetic; D014162:Transfection; D019076:Transgenes; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D050676:Core Binding Factor Alpha 2 Subunit; D037181:Lectins, C-Type; D008156:Luciferases; D000099192:CD69 Antigens; C493728:RUNX1 protein, human
keywords: CD69; CNS; Enhancer; Gene regulation; Transcription regulation