title: GATA2 germline mutations impair GATA2 transcription, causing haploinsufficiency: functional analysis of the p.Arg396Gln mutation.
abstract: Germline GATA2 mutations have been identified as the cause of familial syndromes with immunodeficiency and predisposition to myeloid malignancies. GATA2 mutations appear to cause loss of function of the mutated allele leading to haploinsufficiency; however, this postulate has not been experimentally validated as the basis of these syndromes. We hypothesized that mutations that are translated into abnormal proteins could affect the transcription of GATA2, triggering GATA2 deficiency. Chromatin immunoprecipitation and luciferase assays showed that the human GATA2 protein activates its own transcription through a specific region located at -2.4 kb, whereas the p.Thr354Met, p.Thr355del, and p.Arg396Gln germline mutations impair GATA2 promoter activation. Accordingly, GATA2 expression was decreased to ∼58% in a patient with p.Arg396Gln, compared with controls. p.Arg396Gln is the second most common mutation in these syndromes, and no previous functional analyses have been performed. We therefore analyzed p.Arg396Gln. Our data show that p.Arg396Gln is a loss-of-function mutation affecting DNA-binding ability and, as a consequence, it fails to maintain the immature characteristics of hematopoietic stem and progenitor cells, which could result in defects in this cell compartment. In conclusion, we show that human GATA2 binds to its own promoter, activating its transcription, and that the aforementioned mutations impair the transcription of GATA2. Our results indicate that they can affect other GATA2 target genes, which could partially explain the variability of symptoms in these diseases. Moreover, we show that p.Arg396Gln is a loss-of-function mutation, which is unable to retain the progenitor phenotype in cells where it is expressed.
authors.affiliation: Hematology/Oncology Program, Center for Applied Medical Research, University of Navarra, Pamplona 31008, Spain; Department of Biochemistry and Genetics, University of Navarra, Pamplona 31008, Spain; xcortes@alumni.unav.es.
authors.forename: Xabier
authors.initials: X
authors.lastname: Cortés-Lavaud
authors.affiliation: Department of Internal Medicine, University Clinic of Navarra, Navarra, Pamplona 31008, Spain; and.
authors.forename: Manuel F
authors.initials: MF
authors.lastname: Landecho
authors.affiliation: Hematology/Oncology Program, Center for Applied Medical Research, University of Navarra, Pamplona 31008, Spain;
authors.forename: Miren
authors.initials: M
authors.lastname: Maicas
authors.affiliation: Hematology/Oncology Program, Center for Applied Medical Research, University of Navarra, Pamplona 31008, Spain;
authors.forename: Leire
authors.initials: L
authors.lastname: Urquiza
authors.affiliation: Department of Immunology, University Clinic of Navarra, Pamplona 31008, Spain.
authors.forename: Juana
authors.initials: J
authors.lastname: Merino
authors.affiliation: Hematology/Oncology Program, Center for Applied Medical Research, University of Navarra, Pamplona 31008, Spain;
authors.forename: Isabel
authors.initials: I
authors.lastname: Moreno-Miralles
authors.affiliation: Hematology/Oncology Program, Center for Applied Medical Research, University of Navarra, Pamplona 31008, Spain; Department of Biochemistry and Genetics, University of Navarra, Pamplona 31008, Spain;
authors.forename: María D
authors.initials: MD
authors.lastname: Odero
mesh_terms: D000483:Alleles; D001665:Binding Sites; D045744:Cell Line, Tumor; D005260:Female; D050989:GATA2 Transcription Factor / Q000235:genetics* / Q000276:immunology; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D018095:Germ-Line Mutation; D057895:Haploinsufficiency; D006801:Humans; D007153:Immunologic Deficiency Syndromes / Q000150:complications / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D008875:Middle Aged; D008958:Models, Molecular; D015269:Mycobacterium avium Complex / Q000276:immunology; D015270:Mycobacterium avium-intracellulare Infection / Q000150:complications / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D015398:Signal Transduction; D014158:Transcription, Genetic
chemical_list: D050989:GATA2 Transcription Factor; C494711:GATA2 protein, human