Cloning of a major surface-antigen gene of Trypanosoma cruzi and identification of a nonapeptide repeat.
The parasitic protozoan Trypanosoma cruzi can establish infection in humans and other vertebrate hosts through direct penetration of host cells by trypomastigotes transmitted by the insect vector. Although the molecular processes involved in trypomastigote interiorization of vertebrate cells are unknown, several studies suggest that surface glycoproteins are involved. It is likely that the proteins involved are specific to the trypomastigote stage of the parasite, since only trypomastigotes found in both the insect vector and the vertebrate host bloodstream are capable of invading vertebrate cells. In contrast, the epimastigote stage, found exclusively in the vector, and the amastigote stage, an intracellular stage in the vertebrate host, cannot penetrate the cell directly. We have therefore concentrated our efforts on trypomastigote surface proteins and, along with others, have identified two trypomastigote-specific surface glycoproteins of relative molecular mass (Mr) 90,000 (90K) and 85,000 (85K). Antibody neutralization experiments indicate that the 85K glycoprotein is necessary for efficient interiorization of trypomastigotes in mammalian cells. Here we describe the molecular cloning of a genomic DNA fragment that encodes antigenic determinants present in the 85K trypomastigote surface antigen. The polypeptide fragment encoded by the cloned DNA is recognized by serum from a T. cruzi-infected host and is inferred by DNA sequence analysis to contain a nonapeptide unit that is tandemly repeated five times. Also, the messenger complementary to the cloned DNA fragment is present only in the trypomastigote stage of the parasite.
D000953:Antigens, Protozoan / Q000235:genetics*; D000954:Antigens, Surface / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D000939:Epitopes; D005796:Genes; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis; D014349:Trypanosoma cruzi / Q000235:genetics*
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D000939:Epitopes; D012333:RNA, Messenger