Recombinant and native Plasmodium falciparum TATA-binding-protein binds to a specific TATA box element in promoter regions

Ruvalcaba-Salazar, Omar K.; Ramírez-Estudillo, Ma. del Carmen; Montiel-Condado, Dvorak; Recillas‐Targa, Félix; Vargas, Miguel; Hernández‐Rivas, Rosaura

doi:10.1016/j.molbiopara.2005.01.002

Cited by 36 publications

(29 citation statements)

References 34 publications

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“…Nuclear extract preparation, gel-shift and super-shift assays Preparation of nuclear extracts, gel shift and super-gel shift were performed as described previously (Freitas-Junior et al, 2005;Ruvalcaba-Salazar et al, 2005). Telomere probe of 175 bp was amplified from a plasmid obtained in a previous work (Figueiredo et al, 2000) using the primers listed in supplementary material Table S1.…”

Section: Fluorescence In Situ Hybridizationmentioning

confidence: 99%

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Differential association of Orc1 and Sir2 proteins to telomeric domains in Plasmodium falciparum

Mâncio-Silva

Rojas-Meza

Vargas

et al. 2008

Journal of Cell Science

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Section: Fluorescence In Situ Hybridizationmentioning

confidence: 99%

“…TARE6 probe contains three Rep20 units and was obtained by hybridization of a 79-bp oligonucleotide with the respective anti-sense oligonucleotides (supplementary material Table S1). Kharp and Sp1 probes were obtained as described previously (Ruvalcaba-Salazar et al, 2005).…”

Section: Fluorescence In Situ Hybridizationmentioning

confidence: 99%

Differential association of Orc1 and Sir2 proteins to telomeric domains in Plasmodium falciparum

Mâncio-Silva

Rojas-Meza

Vargas

et al. 2008

Journal of Cell Science

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“…falciparum's steady-state expression patterns during the IDC have been hypothesized to result from cis-acting regulatory DNA sequences (e.g., promoters and enhancers), given its canonical basal transcriptional machinery (4,5,55). Numerous studies of individual putative transcription factors or of individual target genes have found relationships between cis-acting DNA sequences and the expression of individual loci or small groups of related genes (23,38,44,47,53), in some cases demonstrating the activity of putative regulatory proteins on the motifs (18,20,50,62).…”

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confidence: 99%

Patterns of Gene-Specific and Total Transcriptional Activity during thePlasmodium falciparumIntraerythrocytic Developmental Cycle

et al. 2009

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The relationships among gene regulatory mechanisms in the malaria parasite Plasmodium falciparum throughout its asexual intraerythrocytic developmental cycle (IDC) remain poorly understood. To investigate the level and nature of transcriptional activity and its role in controlling gene expression during the IDC, we performed nuclear run-on on whole-transcriptome samples from time points throughout the IDC and found a peak in RNA polymerase II-dependent transcriptional activity related to both the number of nuclei per parasite and variable transcriptional activity per nucleus over time. These differential total transcriptional activity levels allowed the calculation of the absolute transcriptional activities of individual genes from gene-specific nuclear run-on hybridization data. For half of the genes analyzed, sense-strand transcriptional activity peaked at the same time point as total activity. The antisense strands of several genes were substantially transcribed. Comparison of the transcriptional activity of the sense strand of each gene to its steady-state RNA abundance across the time points assayed revealed both correlations and discrepancies, implying transcriptional and posttranscriptional regulation, respectively. Our results demonstrate that such comparisons can effectively indicate gene regulatory mechanisms in P. falciparum and suggest that genes with diverse transcriptional activity levels and patterns combine to produce total transcriptional activity levels tied to parasite development during the IDC.Plasmodium falciparum, the most deadly human malaria parasite, undergoes many developmental changes and thrives in both human and mosquito hosts. Since the genome sequence of P. falciparum became available (17), microarray studies of its intraerythrocytic developmental cycle (IDC) (3, 37) and other asexual and bloodborne stages (34) have demonstrated that differential gene expression is integral to the parasite's development. The stage specificity of steady-state RNA expression supports previous observations that the morphological changes of the IDC correspond to many less-apparent changes in physiology, such as protein and nucleic acid metabolism (10, 19) and sensitivity to small molecules (14, 22, 61). The Plasmodium repertoire of gene regulatory mechanisms is comparable to those of other eukaryotes (7, 23), implying that stage-specific gene expression may be regulated at many levels, including chromatin structure, transcriptional activation, and posttranscriptional modulation of translation and stability (11,39,41,59). However, the relationship between such mechanisms during the IDC, specifically whether regulation of primary transcriptional activity determines the steadystate RNA level, remains uncertain for the vast majority of genes.P. falciparum's steady-state expression patterns during the IDC have been hypothesized to result from cis-acting regulatory DNA sequences (e.g., promoters and enhancers), given its canonical basal transcriptional machinery (4,5,55). Numerous studies of individual puta...

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“…(15) To date, the TATA binding protein (PfTBP) is the only trans-acting factor that has been isolated and its corresponding cis-acting sequences have been identified in two P. falciparum promoters. (2) The TBP protein and many TBP associated factors (TAFs) form the multimeric TFIID complex. Currently, some TAF have been identified (i.e., TAF1, TAF2, TAF7, and TAF5) in P. falciparum.…”

Section: Discussionmentioning

confidence: 99%

“…Currently, a few transcription associated TAPs proteins and general transcription factors have been identified in the P. falciparum genome and unexpected TBP is the only known component of the TFIID complex that has been characterized in this parasite. (2,3) In eukaryotic cells, TBP associates with several subsets of proteins called TATA-binding protein-associated factors (TAFs) that form the multimeric TFIID complex. (4) Analysis of the molecular organization of yeast TFIID (yTFIID) has revealed that more than half (9/14) of the yeast TAFs contain a histone fold motif, and that they specifically assemble into five histone-like pairs.…”

Section: Introductionmentioning

confidence: 99%

Preparation and Characterization of a Monoclonal Antibody Specific to Plasmodium falciparum TATA Binding Protein

et al. 2006

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PfTBP is a transcriptional factor required by all three types of RNA polymerases in eukaryotic cells. In order to obtain a specific monoclonal antibody (MAb) against PfTBP, a DNA fragment of 684 base pairs (bp) that contained the complete PfTBP gene was amplified by polymerase chain reaction (PCR) and inserted into the pGEX prokaryotic expression vector. The recombinant protein (GST-PfTBP) was expressed in Escherichia coli, purified, and used as antigen to immunize mice. MAbs against PfTBP were obtained and hybridomas were screened by enzyme-linked immunosorbent assay (ELISA). Western blotting and immunofluorescence assays showed that MAb Pf.r1 recognized the PfTBP protein in nuclear extracts from Plasmodium falciparum as well as a native protein in the nuclei of this parasite. This MAb will be a helpful tool for the identification of the TBP associated factors (TAFs), which are apparently highly divergent with other eukaryotes. This information could help to identify new candidate gene products to develop novel drugs or vaccines.

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Recombinant and native Plasmodium falciparum TATA-binding-protein binds to a specific TATA box element in promoter regions

Cited by 36 publications

References 34 publications

Differential association of Orc1 and Sir2 proteins to telomeric domains in Plasmodium falciparum

Differential association of Orc1 and Sir2 proteins to telomeric domains in Plasmodium falciparum

Patterns of Gene-Specific and Total Transcriptional Activity during thePlasmodium falciparumIntraerythrocytic Developmental Cycle

Preparation and Characterization of a Monoclonal Antibody Specific to Plasmodium falciparum TATA Binding Protein

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