Abstract
The endosomal compartment performs extensive sorting functions in most eukaryotes, some of which are accomplished with the help of the multivesicular body (MVB) sorting pathway. This pathway depends on the sequential action of complexes, termed the endosomal sorting complex required for transport (ESCRT). After successful sorting, the crucial step of recycling of the ESCRT complex components requires the activation of the AAA ATPase Vps4, and Did2/Vps46 plays an important role in this activation event. The endolysosomal system of the protozoan parasite Giardia lamblia appears to lack complexity, for instead of having distinct early endosomes, late endosomes and lysosomes, there are only peripheral vesicles (PVs) that are located close to the cell periphery. Additionally, comparative genomics studies predict the presence of only a subset of the ESCRT components in G. lamblia. Thus, it is possible that the MVB pathway is not functional in G. lamblia. To address this issue, the present study focused on the two putative orthologues of Did2/Vps46 of G. lamblia as their function is likely to be pivotal for a functional MVB sorting pathway. In spite of considerable sequence divergence, compared to other eukaryotic orthologues, the proteins encoded by both these genes have the ability to function as Did2/Vps46 in the context of the yeast ESCRT pathway. Furthermore, they also localized to the cellular periphery, where PVs are also located. Thus, this report is the first to provide experimental evidence indicating the presence of a functional ESCRT component in G. lamblia by characterizing the putative Did2/Vps46 orthologues.
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Acknowledgments
The authors thank Dr. Alok Kumar Sil for his valuable suggestions during the preparation of this manuscript. This study was supported by funding from Bose Institute. Somnath Dutta is a fellow of the Council of Scientific and Industrial Research (09/015(0423)/2010-EMR-1); Nabanita Saha and Atrayee Ray are fellows of University Grant Commission (F.2-8/2002 (SA-1) dated 04.10.2012 and F.2-8/2002 (SA-1) dated 04.04.2013 respectively). Authors also acknowledge the help of Ms. Boni Halder and DBT-IPLS Confocal Microscopy Facility, University of Calcutta, for assistance with imaging and the Central Instrument Facility of Bose Institute for real-time PCR and DNA sequencing.
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Dutta, S., Saha, N., Ray, A. et al. Significantly Diverged Did2/Vps46 Orthologues from the Protozoan Parasite Giardia lamblia . Curr Microbiol 71, 333–340 (2015). https://doi.org/10.1007/s00284-015-0844-4
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DOI: https://doi.org/10.1007/s00284-015-0844-4