title: Targeted disruption of the CP2 gene, a member of the NTF family of transcription factors.
abstract: The NTF-like family of transcription factors have been implicated in developmental regulation in organisms as diverse as Drosophila and man. The two mammalian members of this family, CP2 (LBP-1c/LSF) and LBP-1a (NF2d9), are highly related proteins sharing an overall amino acid identity of 72%. CP2, the best characterized of these factors, is a ubiquitously expressed 66-kDa protein that binds the regulatory regions of many diverse genes. Consequently, a role for CP2 has been proposed in globin gene expression, T-cell responses to mitogenic stimulation, and several other cellular processes. To elucidate the in vivo role of CP2, we have generated mice nullizygous for the CP2 allele. These animals were born in a normal Mendelian distribution and displayed no defects in growth, behavior, fertility, or development. Specifically, no perturbation of hematopoietic differentiation, globin gene expression, or immunological responses to T- and B-cell mitogenic stimulation was observed. RNA and protein analysis confirmed that the nullizygous mice expressed no full-length or truncated version of CP2. Electrophoretic mobility shift assays with nuclear extracts from multiple tissues demonstrated loss of CP2 DNA binding activity in the -/- lines. However, a slower migrating complex that was ablated with antiserum to NF2d9, the murine homologue of LBP-1a, was observed with these extracts. Furthermore, we demonstrate that recombinant LBP-1a can bind to known CP2 consensus sites and form protein complexes with previously defined heteromeric partners of CP2. These results suggest that LBP-1a/NF2d9 may compensate for loss of CP2 expression in vivo and that further analysis of the role of the NTF family of proteins requires the targeting of the NF2d9 gene.
authors.affiliation: Division of Experimental Hematology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
authors.forename: L
authors.initials: L
authors.lastname: Ramamurthy
authors.forename: V
authors.initials: V
authors.lastname: Barbour
authors.forename: A
authors.initials: A
authors.lastname: Tuckfield
authors.forename: D R
authors.initials: DR
authors.lastname: Clouston
authors.forename: D
authors.initials: D
authors.lastname: Topham
authors.forename: J M
authors.initials: JM
authors.lastname: Cunningham
authors.forename: S M
authors.initials: SM
authors.lastname: Jane
mesh_terms: D000818:Animals; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics*; D004622:Embryo, Mammalian / Q000378:metabolism; D018390:Gene Targeting; D006410:Hematopoiesis; D051379:Mice; D008822:Mice, Transgenic; D016601:RNA-Binding Proteins; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D016601:RNA-Binding Proteins; C501905:Tfcp2 protein, mouse; D014157:Transcription Factors; C094751:Ubp1 protein, mouse; D004247:DNA