TY - JOUR
T1 - Analysis of the Leishmania donovani transcriptome reveals an ordered progression of transient and permanent changes in gene expression during differentiation
AU - Saxena, A.
AU - Lahav, T.
AU - Holland, N.
AU - Aggarwal, G.
AU - Anupama, A.
AU - Huang, Y.
AU - Volpin, H.
AU - Myler, P. J.
AU - Zilberstein, D.
N1 - Funding Information:
We thank Dr. Stephen Beverley (Washington University) for the kind gift of the LmjF GSS clones and end-sequences (GenBank Accession numbers AQ843743 – AQ853356 , AQ901732 – AQ902705 and AQ911373 – AQ912039 ), the SBRI genome sequencing team for additional end-sequencing, Aaron Leland for his technical assistance with RNA labeling, and Dr. Deborah Smith for her gift of the pDA-PET 33/7 plasmid. This work was supported by PHS grant AI47234 to PJM from the National Institutes of Health, Grant T24-86-1 from the UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases and grant 2003237 from the U.S.-Israel Binational Foundation.
PY - 2007/3
Y1 - 2007/3
N2 - Leishmania donovani is an intracellular protozoan parasite that causes kala-azar in humans. During infection the extracellular insect forms (promastigotes) undergo rapid differentiation to intracellular amastigotes that proliferates in phagolysosomes of mammalian macrophages. We used microarray-based expression profiling to investigate the time-course of changes in RNA abundance during promastigote-to-amastigote differentiation in a host-free system that mimics this process. These studies revealed that several hundred genes underwent an ordered progression of transient or permanent up- and down-regulation during differentiation. Genes that were permanently up-regulated in amastigotes were enriched for transporters and surface proteins, but under-represented in genes involved in protein and other metabolism. Most of these changes occurred late in the differentiation process, when morphological differentiation was essentially complete. Down-regulated genes were over-represented in those involved in cell motility, growth and/or maintenance, and these changes generally occurred earlier in the process. Genes that were transiently up- or down-regulated during differentiation included those encoding heat shock proteins, ubiquitin hydrolases, RNA binding proteins, protein kinases, a protein phosphatase, and a histone deacetylase. These results suggest that changes in mRNA abundance may be important in signal transduction, as well as protein and mRNA turnover, during differentiation. In addition to these mRNA changes, other transcripts including one or more rRNAs and snoRNAs, and non-coding RNAs from several telomeres, also showed substantial changes in abundance during the differentiation process. This paper provides the first genome-scale quantitative analysis of gene expression during the transition from promastigotes to amastigotes and demonstrates the utility of the host-free differentiation system.
AB - Leishmania donovani is an intracellular protozoan parasite that causes kala-azar in humans. During infection the extracellular insect forms (promastigotes) undergo rapid differentiation to intracellular amastigotes that proliferates in phagolysosomes of mammalian macrophages. We used microarray-based expression profiling to investigate the time-course of changes in RNA abundance during promastigote-to-amastigote differentiation in a host-free system that mimics this process. These studies revealed that several hundred genes underwent an ordered progression of transient or permanent up- and down-regulation during differentiation. Genes that were permanently up-regulated in amastigotes were enriched for transporters and surface proteins, but under-represented in genes involved in protein and other metabolism. Most of these changes occurred late in the differentiation process, when morphological differentiation was essentially complete. Down-regulated genes were over-represented in those involved in cell motility, growth and/or maintenance, and these changes generally occurred earlier in the process. Genes that were transiently up- or down-regulated during differentiation included those encoding heat shock proteins, ubiquitin hydrolases, RNA binding proteins, protein kinases, a protein phosphatase, and a histone deacetylase. These results suggest that changes in mRNA abundance may be important in signal transduction, as well as protein and mRNA turnover, during differentiation. In addition to these mRNA changes, other transcripts including one or more rRNAs and snoRNAs, and non-coding RNAs from several telomeres, also showed substantial changes in abundance during the differentiation process. This paper provides the first genome-scale quantitative analysis of gene expression during the transition from promastigotes to amastigotes and demonstrates the utility of the host-free differentiation system.
KW - Axenic amastigotes
KW - Differentiation
KW - Gene expression
KW - L. donovani
KW - Transcript abundance
UR - http://www.scopus.com/inward/record.url?scp=33846259286&partnerID=8YFLogxK
U2 - 10.1016/j.molbiopara.2006.11.011
DO - 10.1016/j.molbiopara.2006.11.011
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C2 - 17204342
AN - SCOPUS:33846259286
SN - 0166-6851
VL - 152
SP - 53
EP - 65
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology
IS - 1
ER -