Expression of trans-sialidase and 85-kDa glycoprotein genes in Trypanosoma cruzi is differentially regulated at the post-transcriptional level by labile protein factors.
To adapt to different environments, Trypanosoma cruzi, the protozoan parasite that causes Chagas' disease, expresses a different set of proteins during development. To begin to understand the mechanism that controls this differential gene expression, we have analyzed the levels of amastin and trans-sialidase mRNAs and the mRNAs encoding members of the 85-kDa glycoprotein gene family, which are differentially expressed in the T. cruzi stages found in the mammalian host. Amastin mRNA is expressed predominantly in intracellular and proliferative amastigotes. trans-Sialidase mRNAs are found mostly in forms undergoing transformation from amastigotes to trypomastigotes inside infected cells, whereas mRNAs encoding the 85-kDa glycoproteins appear only in the infective trypomastigotes released from the cells. The genes coding for these mRNA species are constitutively transcribed in all stages of T. cruzi cells, suggesting that expression is controlled post-transcriptionally during differentiation. Inhibition of transcription by actinomycin D revealed that each mRNA species has a relatively long half-life in stages where it accumulates. In the case of the trans-sialidase and 85-kDa glycoprotein genes, mRNA accumulation was induced by treatment with the protein synthesis inhibitor cycloheximide at the stages that preceded the normal accumulation. Therefore, mRNA stabilization may account for mRNA accumulation. mRNA degradation could be promoted by proteins with high turnover, or stabilization could be promoted by forming a complex with the translational machinery at defined times in development. Identification of the factors that induce mRNA degradation or stabilization is essential to the understanding of control of gene expression in these organisms.
Abuin
G
G
Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, R. Botucatu 862 8o A, 04023-062 São Paulo, São Paulo, Brasil.
Freitas-Junior
L H
LH
Colli
W
W
Alves
M J
MJ
Schenkman
S
S
D000818:Animals; D001483:Base Sequence; D003513:Cycloheximide / Q000494:pharmacology; D017931:DNA Primers; D003609:Dactinomycin / Q000494:pharmacology; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000235:genetics*; D009439:Neuraminidase / Q000235:genetics*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014349:Trypanosoma cruzi / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D006023:Glycoproteins; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D016053:RNA, Protozoan; D003609:Dactinomycin; D003513:Cycloheximide; D009439:Neuraminidase