Hyperosmotic stress induces aquaporin-dependent cell shrinkage, polyphosphate synthesis, amino acid accumulation, and global gene expression changes in Trypanosoma cruzi.
The protist parasite Trypanosoma cruzi has evolved the ability to transit between completely different hosts and to replicate in adverse environments. In particular, the epimastigote form, the replicative stage inside the vector, is subjected to nutritional and osmotic stresses during its development. In this work, we describe the biochemical and global gene expression changes of epimastigotes under hyperosmotic conditions. Hyperosmotic stress resulted in cell shrinking within a few minutes. Depending on the medium osmolarity, this was followed by lack of volume recovery for at least 2 h or by slow recovery. Experiments with inhibitors, or with cells in which an aquaporin gene (TcAQP1) was knocked down or overexpressed, revealed its importance for the cellular response to hyperosmotic stress. Furthermore, the adaptation to this new environment was shown to involve the regulation of the polyphosphate polymerization state as well as changes in amino acid catabolism to generate compatible osmolytes. A genome-wide transcriptional analysis of stressed parasites revealed down-regulation of genes belonging to diverse functional categories and up-regulation of genes encoding trans-sialidase-like and ribosomal proteins. Several of these changes were confirmed by Northern blot analyses. Sequence analysis of the 3'UTRs of up- and down-regulated genes allowed the identification of conserved structural RNA motifs enriched in each group, suggesting that specific ribonucleoprotein complexes could be of great importance in the adaptation of this parasite to different environments through regulation of transcript abundance.
Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia 30602.
Zhu-Hong
ZH
Li
Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia 30602; Instituto de Investigaciones Biotecnólogicas, Universidad Nacional de General San Martin/Consejo Nacional de Investigaciones Científicas y Técnicas, 1650 San Martin, Buenos Aires, Argentina.
Vanina E
VE
Alvarez
Instituto de Investigaciones Biotecnólogicas, Universidad Nacional de General San Martin/Consejo Nacional de Investigaciones Científicas y Técnicas, 1650 San Martin, Buenos Aires, Argentina.
Javier G
JG
De Gaudenzi
Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia 30602.
Celso
C
Sant'Anna
Instituto de Investigaciones Biotecnólogicas, Universidad Nacional de General San Martin/Consejo Nacional de Investigaciones Científicas y Técnicas, 1650 San Martin, Buenos Aires, Argentina.
Alberto C C
ACC
Frasch
Instituto de Investigaciones Biotecnólogicas, Universidad Nacional de General San Martin/Consejo Nacional de Investigaciones Científicas y Técnicas, 1650 San Martin, Buenos Aires, Argentina.
Juan J
JJ
Cazzulo
Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, Georgia 30602. Electronic address: rdocampo@uga.edu.
Roberto
R
Docampo
D000596:Amino Acids / Q000737:chemistry*; D000818:Animals; D020346:Aquaporins / Q000737:chemistry*; D002462:Cell Membrane / Q000378:metabolism; D015536:Down-Regulation; D015870:Gene Expression; D005786:Gene Expression Regulation; D008854:Microscopy, Electron / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D009995:Osmosis; D011122:Polyphosphates / Q000737:chemistry*; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014349:Trypanosoma cruzi / Q000378:metabolism*; D015854:Up-Regulation
D000596:Amino Acids; D020346:Aquaporins; D011122:Polyphosphates; D012333:RNA, Messenger