Recombinant and native Plasmodium falciparum TATA-binding-protein binds to a specific TATA box element in promoter regions.
RNA polymerase II promoters in Plasmodium spp., like in most eukaryotes, have a bipartite structure. However, the identification of a functional TATA box located within the Plasmodium spp. core promoters has been difficult, mainly because of its high A+T content. Only few putative trans-acting elements have been identified in the malaria parasite genome such as a gene orthologous to the TATA box binding protein (PfTBP). In this study, we demonstrate that PfTBP is part of the DNA-protein complexes formed in the kahrp and gbp-130 gene promoter regions. Supershift and footprinting assays performed with a GST-PfTBP fusion protein showed that PfTBP associates with a consensus TATA box sequence located 81 base pairs upstream of the transcription start site in the kahrp promoter region and with a TATA box-like (TGTAA) sequence at position -186 of the gbp-130 gene promoter region. Chromatin immunoprecipitation assays confirmed that native PfTBP is able to associate in vivo with both TATA box elements. This is the first study that reports the identification of cis-acting sequences (TATAA and TGTAA) and their corresponding trans-acting (PfTBP) factor in P. falciparum.
Ruvalcaba-Salazar
Omar K
OK
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (IPN), Apartado Postal 14-740, 07360 México.
del Carmen Ramírez-Estudillo
Ma
M
Montiel-Condado
Dvorak
D
Recillas-Targa
Félix
F
Vargas
Miguel
M
Hernández-Rivas
Rosaura
R
D000818:Animals; D001665:Binding Sites; D010455:Peptides / Q000235:genetics; D010963:Plasmodium falciparum / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D015800:Protozoan Proteins / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D016385:TATA Box; D035181:TATA-Box Binding Protein / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D024363:Transcription Initiation Site
D010455:Peptides; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D035181:TATA-Box Binding Protein; C042968:knob protein, Plasmodium falciparum; C072465:glycophorin-binding protein homologue, Plasmodium