Upstream regulatory elements controlling expression of the Entamoeba histolytica lectin

Purdy, Jay E.; Pho, Lana T.; Mann, Barbara J.; Petri, William A.

doi:10.1016/s0166-6851(96)02614-x

Cited by 93 publications

(104 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Many genes shown to possess both a TATA-(or TATA-like) box and an initiator are such that at least one of these two motifs is sufficient to drive substantial transcription if either motif is mutationally disabled (e.g. [34,38,[41][42][43][44]). We also observed that in cell lines, transcription was essentially abolished when the initiator was debilitated and it was decreased when the ATATAT motif was removed.…”

Section: Discussionmentioning

confidence: 99%

Transcription of the juvenile hormone esterase gene under the control of both an initiator and AT-rich motif

Jones

Mańczak

Schelling

et al. 1998

Biochemical Journal

View full text Add to dashboard Cite

The binding of transcription factors to the core promoter of the juvenile hormone esterase gene was functionally characterized using both a cell-free in vitrotranscription functional assay and a cell transfection assay. A core JHE promoter (-61 to +28 bp relative to transcription start site) supported faithful transcription from the in vivo transcription start site. The nuclear extracts from the Sf9 insect cell line that provided transcription from that template also bound to that template as a probe in gel-mobility shift assays. Deletion or transversion of the initiator-binding motif (-1 to +4 bp) abolished detectable transcription either in vitro or in transfected cells. An AT-rich motif (ATATAT; -28 to -23 bp) serves another transcription factor-binding site. Mutation of the AT-rich motif to a canonical TATA-box preserved transcription, while either its deletion or complete transversion abolished or significantly reduced detectable transcriptional activity. These results indicate that, under these conditions, the functional operation of this core promoter approaches that of a composite promoter in which both the TATA- and initiator-binding protein complexes are necessary, even for basal transcription. On the other hand, these debilitating mutations to either the TATA box or initiator motif did not prevent the ability of the corresponding gel-shift competitive probes to compete with the wild-type promoter for binding by the transcription factors. Even a double transversion of both the AT-rich motif and the initiator-binding motif was able to competitively displace the protein complex that bound to the labelled wild-type probe. These data strongly indicate the presence of (an) additional core-promoter-associated transcription factor(s) (that is not the ‘downstream element ’) that contact(s) the AT-binding complex and/or initiator-binding factor with sufficient avidity to remove them from binding to the competing wild-type promoter sequence.

show abstract

Section: Discussionmentioning

confidence: 99%

Transcription of the juvenile hormone esterase gene under the control of both an initiator and AT-rich motif

Jones

Mańczak

Schelling

et al. 1998

Biochemical Journal

View full text Add to dashboard Cite

show abstract

“…Construction of the pTP.4i plasmid was described earlier and consists of a 272-bp hgl5 upstream region fused to a luciferase reporter gene and the 5Ј actin region fused to the neomycin drug selection gene (9,17,20). In the plasmid pTP.GAAC-CLA the 5Ј core promoter region of the hgl5 gene has the GAAC region mutated to a ClaI site (GAACT to CGATT).…”

Section: Cultivation Of E Histolytica and Stablementioning

confidence: 99%

“…In the plasmid pTP.GAAC-CLA the 5Ј core promoter region of the hgl5 gene has the GAAC region mutated to a ClaI site (GAACT to CGATT). To introduce a GAAC region in the upstream location, we used a two-round PCR technique (17). Previous analysis of the 5Ј-noncoding region of the hgl5 promoter had revealed that the region 20 bases upstream of the TATA sequence could be replaced by an AT-rich sequence without affecting gene expression (17).…”

Section: Cultivation Of E Histolytica and Stablementioning

confidence: 99%

“…The amebic TATA (GTATTTAAA(G/C)) and Inr (AAAAATTCA) elements appear to function in a classic manner despite their sequence divergence from metazoans (9). A unique third core promoter element GAAC (GAACT) was identified in 31/37 protein-encoding E. histolytica genes (17). It has a variable location between the TATA and Inr sequences, a characteristic not defined previously for core promoter transcription elements in metazoans.…”

mentioning

confidence: 93%

See 1 more Smart Citation

The Novel Core Promoter Element GAAC in the hgl5 Gene of Entamoeba histolytica Is Able to Direct a Transcription Start Site Independent of TATA or Initiator Regions

Singh

Rogers

1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

Entamoeba histolytica, an enteric protozoa, is the third leading parasitic cause of death worldwide. Investigation of the transcriptional machinery of this eukaryotic pathogen has revealed an unusual core promoter structure that consists of nonconsensus TATA and initiator regions and a novel third conserved core promoter sequence, the GAAC element. Mutation of this region in the hgl5 promoter decreases reporter gene expression and alters the transcription start site. Using positional analysis of this element, we have now demonstrated that it is able to direct a new transcription start site, 2-7 bases downstream of itself, independent of TATA and Inr regions. The GAAC region was also shown to control the rate of transcription via nuclear run on analysis and an amebic nuclear protein was demonstrated to specifically interact with this sequence. This is the first description in the eukaryotic literature of a third conserved core promoter element, distinct from TATA or initiator regions, that is able to direct a transcription start site. We have formulated two models for the role of the GAAC region: (i) the GAAC-binding protein is a part of the TFIID complex and (ii) the GAAC-binding protein functions to "tether" TATA-binding protein to the TATA box.

show abstract

“…Just four promoters from the EhPgp1, EhPgp5, hgl5 and actin genes have been functionally characterized and until this moment have been functionally described the TATA box, the CAAT/enhancer binding elements, the GAACT and the UREs sequences [16][17][18][19][20][21][22][23][24]. With respect to the transcription factors have been reported the sequences for the TATA box binding protein (TBP) [25], two enhancer binding proteins (EhEBP1 and EhEBP2) that binds to the URE4 sequence [26] and the p53 [27] and C/EBP-like proteins (personal communication).…”

Section: Introductionmentioning

confidence: 99%

A Novel Heat Shock Transcription Factor Family in Entamoeba histolytica

García¹,

Argüelles²,

Ishiwara³

et al. 2007

American J. of Infectious Diseases

View full text Add to dashboard Cite

Abstract:The HSTF is a master molecule involved in the transcriptional control of several genes during different types of stress. This transcription factor is a very conserved protein identified in different organisms from bacterial to human. Entamoeba histolytica is the protozoan responsible for the human amoebiasis. This parasite is exposed to different kind of stress as changes in the pH, temperature, drugs, all that situations in where the parasite needs survive. Here we identified and isolated a novel gene family of HSTFs in the protozoan parasite E. histolytica. Three members that we called Ehhstf1, Ehhstf2 and Ehhstf3 compose this family. Amino acid alignments and domain architecture analysis revealed that the EhHSTFs presents a conserved DNA-binding domain composed of approximately 25 residues. Interestingly this domain is shorter than the domain of the human, mouse and yeast HSTFs. Heterologous antibodies recognized four peptides of 73, 66, 47 and 23 kDa in total extracts from trophozoites growth under normal conditions. The 73, 47 and 23 kDa peptides increased their intensity when the cells were growth at 42°C by 2 h. All results together demonstrate that the amoeba present HSTFs, which may be, controlled the gene expression of this parasite under different stress situations.

show abstract

Upstream regulatory elements controlling expression of the Entamoeba histolytica lectin

Cited by 93 publications

References 26 publications

Transcription of the juvenile hormone esterase gene under the control of both an initiator and AT-rich motif

Transcription of the juvenile hormone esterase gene under the control of both an initiator and AT-rich motif

The Novel Core Promoter Element GAAC in the hgl5 Gene of Entamoeba histolytica Is Able to Direct a Transcription Start Site Independent of TATA or Initiator Regions

A Novel Heat Shock Transcription Factor Family in Entamoeba histolytica

Contact Info

Product

Resources

About