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Eukaryotic Cell, March 2009, p. 327-338, Vol. 8, No. 3
1535-9778/09/$08.00+0     doi:10.1128/EC.00340-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Patterns of Gene-Specific and Total Transcriptional Activity during the Plasmodium falciparum Intraerythrocytic Developmental Cycle {triangledown} ,{dagger}

Jennifer S. Sims,1,2 Kevin T. Militello,3 Peter A. Sims,4 Vishal P. Patel,1,5 Jacob M. Kasper,1 and Dyann F. Wirth1,2*

Department of Immunology and Infectious Diseases, Harvard School of Public Health, Harvard University, Boston, Massachusetts 02115,1 Department of Microbiology and Molecular Genetics, Harvard Medical School, 240 Longwood Avenue, Boston, Massachusetts 02115,2 Department of Biology, State University of New York at Geneseo, 1 College Circle, Geneseo, New York 14454,3 Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138,4 Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 021155

Received 9 October 2008/ Accepted 30 December 2008

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.

* Corresponding author. Mailing address: Harvard School of Public Health, Dept. of Immunology and Infectious Disease, Boston, MA 02115. Phone: (617) 432-4629. Fax: (617) 432-4766. E-mail: dfwirth{at}hsph.harvard.edu

{triangledown} Published ahead of print on 16 January 2009.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, March 2009, p. 327-338, Vol. 8, No. 3
1535-9778/09/$08.00+0     doi:10.1128/EC.00340-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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