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Titolo:
In vivo analysis of retroviral enhancer mutations in hematopoietic cells: SP1/EGR1 and ETS/GATA motifs contribute to long terminal repeat specificity
Autore:
Wahlers, A; Zipfel, PF; Schwieger, M; Ostertag, W; Baum, C;
Indirizzi:
Hannover Med Sch, Dept Hematol & Oncol, D-30625 Hannover, Germany HannoverMed Sch Hannover Germany D-30625 col, D-30625 Hannover, Germany Heinrich Pette Inst Expt Virol & Immunol, Dept Cell & Virus Genet, D-20251Hamburg, Germany Heinrich Pette Inst Expt Virol & Immunol Hamburg GermanyD-20251 Germany Kans Knoell Inst Nat Prod Res, D-07745 Jena, Germany Kans Knoell Inst Nat Prod Res Jena Germany D-07745 D-07745 Jena, Germany
Titolo Testata:
JOURNAL OF VIROLOGY
fascicolo: 1, volume: 76, anno: 2002,
pagine: 303 - 312
SICI:
0022-538X(200201)76:1<303:IVAORE>2.0.ZU;2-8
Fonte:
ISI
Lingua:
ENG
Soggetto:
MURINE LEUKEMIA-VIRUS; FOCUS-FORMING VIRUS; MYELOPROLIFERATIVE SARCOMA-VIRUS; MARROW TRANSPLANT RECIPIENTS; GREEN FLUORESCENT PROTEIN; STEM-CELLS; CONTROL REGION; BINDING-SITES; BONE-MARROW; TRANSCRIPTIONAL ACTIVATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
54
Recensione:
Indirizzi per estratti:
Indirizzo: Baum, C Hannover Med Sch, Dept Hematol & Oncol, OE 5120 Carl Neuberg Str 1, D-30625 Hannover, Germany Hannover Med Sch OE 5120 Carl Neuberg Str 1 Hannover Germany D-30625
Citazione:
A. Wahlers et al., "In vivo analysis of retroviral enhancer mutations in hematopoietic cells: SP1/EGR1 and ETS/GATA motifs contribute to long terminal repeat specificity", J VIROLOGY, 76(1), 2002, pp. 303-312

Abstract

The objective of this work was to identify, in the context of chromosomally integrated DNA, the contribution of defined transcription factor binding motifs to the function of a complex retrovirus enhancer in hematopoietic cells in vivo. Repopulating murine hematopoietic cells were transduced with equal gene dosages of replication-incompetent retrovirus vectors encoding enhanced green fluorescent protein. Enhancer sequences were derived from mouse spleen focus-forming virus. Destruction of GC-rich sites representing overlapping targets for SP1 or EGR1 uniformly attenuated gene expression (similar to 25 to 70% of wild-type levels) in all hematopoietic lineages, as shown by multicolor flow cytometry of peripheral blood and bone marrow cells at various time points posttransplantation. In contrast, a point mutation within a dual ETS/GATA motif that abolished transactivation by ETS factors but not by GATA-1 slightly increased activity in erythroid cells and significantly attenuated enhancer function in T lymphocytes. This study shows that controlled gene transfer in transplantable hematopoietic cells allows a functional analysis of distinct cis elements within a complex retrovirus enhancer, as required for the characterization and engineering of various cellular and viral regulatory sequences in basic research and gene therapy.

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Documento generato il 27/01/20 alle ore 07:30:15