Hehl, A. B., Marti, M. and Köhler, P. (2000) Stage-specific expression and targeting of cyst wall protein-green fluorescent protein chimeras in Giardia. Molecular Biology of the Cell, 11(5), pp. 1789-1800. (doi: 10.1091/mbc.11.5.1789) (PMID:10793152) (PMCID:PMC14884)
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Abstract
In preparation for being shed into the environment as infectious cysts, trophozoites of Giardia spp. synthesize and deposit large amounts of extracellular matrix into a resistant extracellular cyst wall. Functional aspects of this developmentally regulated process were investigated by expressing a series of chimeric cyst wall protein 1 (CWP1)–green fluorescent protein (GFP) reporter proteins. It was demonstrated that a short 110 bp 5′ flanking region of the CWP1 gene harbors all necessary cis-DNA elements for strictly encystation-specific expression of a reporter during in vitro encystation, whereas sequences in the 3′ flanking region are involved in modulation of steady-state levels of its mRNA during encystation. Encysting Giardia expressing CWP1–GFP chimeras showed formation and maturation of labeled dense granule-like vesicles and subsequent incorporation of GFP-tagged protein into the cyst wall, dependent on which domains of CWP1 were included. The N-terminal domain of CWP1 was required for targeting GFP to regulated compartments of the secretory apparatus, whereas a central domain containing leucine-rich repeats mediated association of the chimera with the extracellular cyst wall. We show that analysis of protein transport using GFP-tagged molecules is feasible in an anaerobic organism and provides a useful tool for investigating the organization of primitive eukaryotic vesicular transport.
Item Type: | Articles |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Marti, Professor Matthias |
Authors: | Hehl, A. B., Marti, M., and Köhler, P. |
College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity |
Journal Name: | Molecular Biology of the Cell |
Publisher: | American Society for Cell Biology |
ISSN: | 1059-1524 |
ISSN (Online): | 1939-4586 |
Copyright Holders: | Copyright © 2000 The American Society for Cell Biology |
First Published: | First published in Molecular Biology of the Cell 11(5):1789-1800 |
Publisher Policy: | Reproduced under a Creative Commons License |
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