Positive and negative effects of deletions and mutations within the 5' flanking sequences of Plasmodium falciparum DNA polymerase δ

Abstract The DNA polymerase δ gene from Plasmodium falciparum is associated with a 1.76 kb 5′ untranslated region and a promoter containing long homopolymeric (dA:dT) tracts and intrinsic DNA curvature. Here, a comprehensive deletion and mutational analysis of the DNA Polδ upstream sequences has been undertaken to define functionally important regions. Removal of promoter-proximal DNA was shown to upregulate luciferase reporter gene expression and the ATG-proximal portion of the 5′ untranslated region was required in conjunction with the promoter for reporter activity. In contrast to the ATG-proximal 5′ untranslated region, deletion of the central part of the untranslated region had a positive effect on expression. Disruption of a homopolymeric (dA:dT) tract adjacent to the main transcription start site both derepressed gene expression and reduced the intrinsic curvature of DNA fragments containing this sequence.

[1]  Eukaryotic DNA polymerases. , 1991, Annual review of biochemistry.

[2]  J. White,et al.  DNA replication in the malaria parasite. , 1996, Parasitology today.

[3]  V. de Lorenzo,et al.  Clues and consequences of DNA bending in transcription. , 1997, Annual review of microbiology.

[4]  S K Burley,et al.  TATA element recognition by the TATA box-binding protein has been conserved throughout evolution. , 1999, Genes & development.

[5]  W. Trager,et al.  Cultivation of malarial parasites , 1978, Nature.

[6]  D. Battistutta,et al.  Analysis of the sequences flanking the translational start sites of Plasmodium falciparum. , 1990, Molecular and biochemical parasitology.

[7]  J. White,et al.  Stage specific expression of proliferating cell nuclear antigen and DNA polymerase delta from Plasmodium falciparum. , 1996, Molecular and biochemical parasitology.

[8]  Koen J. Dechering,et al.  Isolation and Functional Characterization of Two Distinct Sexual-Stage-Specific Promoters of the Human Malaria Parasite Plasmodium falciparum , 1999, Molecular and Cellular Biology.

[9]  R. Carter,et al.  Plasmodium falciparum: parasites defective in early stages of gametocytogenesis. , 1995, Experimental parasitology.

[10]  M. Porter.  The DNA polymerase delta promoter from Plasmodium falciparum contains an unusually long 5' untranslated region and intrinsic DNA curvature. , 2001, Molecular and biochemical parasitology.

[11]  J. Weber,et al.  Molecular biology of malaria parasites. , 1988, Experimental parasitology.

[12]  M Lanzer,et al.  Mutational analysis identifies a five base pair cis-acting sequence essential for GBP130 promoter activity in Plasmodium falciparum. , 1999, Molecular and biochemical parasitology.

[13]  A. Macleod,et al.  Plasmodium falciparum: modifications of the in vitro culture conditions improving parasitic yields. , 1982, The Journal of parasitology.

[14]  J. Daily,et al.  Deletion analysis of the 5' flanking sequence of the Plasmodium gallinaceum sexual stage specific gene pgs28 suggests a bipartite arrangement of cis-control elements. , 2001, Molecular and biochemical parasitology.

[15]  Efficient extraction and translation of Plasmodium falciparum messenger RNA. , 1982, Molecular and biochemical parasitology.

[16]  S. Jain,et al.  DNA polymerases alpha, delta, and epsilon: three distinct enzymes from HeLa cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Y. Wu.  Transfection of Plasmodiium falciparum within human red biood cells , 1995 .

[18]  H. D. del Portillo,et al.  Comparison of introns in a cdc2-homologous gene within a number of Plasmodium species. , 1995, Molecular and biochemical parasitology.

[19]  B. Stillman,et al.  Replication factors required for SV40 DNA replication in vitro. II. Switching of DNA polymerase alpha and delta during initiation of leading and lagging strand synthesis. , 1991, The Journal of biological chemistry.

[20]  B. Kilbey,et al.  Physical and functional mapping of the transcriptional start sites of Plasmodium falciparum proliferating cell nuclear antigen. , 1996, Molecular and biochemical parasitology.

[21]  J. Ito,et al.  Compilation, alignment, and phylogenetic relationships of DNA polymerases. , 1993, Nucleic acids research.

[22]  X. Su,et al.  Transfection of Plasmodium falciparum within human red blood cells. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[23]  C. Lambros,et al.  Synchronization of Plasmodium falciparum erythrocytic stages in culture. , 1979, The Journal of parasitology.

[24]  John H. White,et al.  DNA polymerase δ: gene sequences from Plasmodium falciparum indicate that this enzyme is more highly conserved than DNA polymerase α , 1991 .