Accumulation of tropomyosin isoform 5 at the infection sites of host cells during Cryptosporidium invasion.
The actin cytoskeleton of host cells has been implicated in Cryptosporidium invasion. However, the underlying mechanism of how actin filaments and associated proteins modulate this process remains unclear. In this study, we use in vitro cultured cell lines, human ileocecal adenocarcinoma HCT-8 and Chinese hamster ovary (CHO), and an in vivo mouse model to investigate the roles of tropomyosin isoforms in Cryptosporidium invasion. Using isoform-specific monoclonal antibodies, we found that the major human tropomyosin (hTM) isoforms expressed in HCT-8 cells are hTM4 and hTM5. HCT-8 cells also express hTM1 at low levels but not hTM2 and hTM3. During Cryptosporidium parvum infection, hTM5 colocalized to the infection sites with a novel parasite membrane protein, CP2. Neither hTM1 nor hTM4 accumulated at infection sites. Similarly, a high level of TM5 and varying amounts of TM4 accumulated at the C. parvum infection sites in CHO cells. CHO cells overexpressing hTM5 exhibit a significantly higher percent of mature meronts early in the infection process relative to CHO cells or CHO cells overexpressing a tropomyosin mutant, chimeric isoform hTM5/3. These results suggest that functional TM5 enhances Cryptosporidium invasion of host cells. In C. parvum-infected mice, accumulation and rearrangement of TM5 and TM4 were detected throughout the infected ileum. Similarly, in the Cryptosporidium muris-infected mice, TM5 accumulated in discrete regions of the epithelial cells of gastric glands and in the oocyst-laden stomach gland lumen. Cryptosporidium infection appears to rearrange and recruit host TM isoforms in both culture cells and in the mouse. Localized accumulation of tropomyosin at the infection sites may facilitate parasite invasion.
O'Hara
Steven P
SP
Department of Biological Sciences, University of Iowa, Iowa City, IA 52242-1324, USA.
Lin
Jim Jung-Ching
JJ
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003457:Cryptosporidiosis / Q000378:metabolism*; D003458:Cryptosporidium; D005260:Female; D005455:Fluorescent Antibody Technique; D005753:Gastric Mucosa / Q000378:metabolism; D006801:Humans; D007082:Ileum / Q000378:metabolism; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D020033:Protein Isoforms / Q000378:metabolism; D015800:Protozoan Proteins / Q000378:metabolism*; D014335:Tropomyosin / Q000378:metabolism*
D008565:Membrane Proteins; D020033:Protein Isoforms; D015800:Protozoan Proteins; C498783:TPM3 protein, human; D014335:Tropomyosin