cells, GlVps might dissociate from AcPh when they reach the acidic pH of the PVs and the receptor recycles back via the retromer complex. In fact, the cargo-selection subunit GlVps35, homologous to Vps35 of the yeast retromer, was pulled-down together with AcPh and GlVps. Since there is no distinguished prevacuolar compartment in Giardia, it is possible that this process of docking-dissociation-recycling occurs almost at the same time, with the AcPh-GlVps-GlVps35 interaction being possible. Experiments concentrating on the functional participation of GlVps35 and the Giardia retromer complex in GlVps recycling are currently underway to test this hypothesis. Also, a combination of genetic and biochemical approaches analyzing the interaction between AcPh and kinesin might contribute to the elucidation of the link between lysosomal protein trafficking and microtubule tracks, where the kinesin-like motor protein might be involved. The identification of the WD40 domain in GlVps together with its subcellular localization suggest that it might behave like the human WD40 protein, WIPI49 or the yeastorthologous Vps18p, with the capacity to regulate endosomal trafficking of proteins and autophagosome formation. Besides the presence of the immunoreactive band at the size of GlVps-HA, 
a higher band, probably PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22212565 corresponding to the size of GlVps homodimer, was observed, suggesting that this protein might interact with itself, as has been shown for several proteins containing one or more WD40 domains. However, we cannot exclude the possibility that the higher molecular weight complex also contains a different protein of similar size that interacts with GlVps. The absence of a cleavable N-terminal signal peptide, and the use of a signal-anchor sequence that directs translocation of the N-terminal domain across the membrane, designate GlVps as a type III-like membrane protein. In the case of GlVps, the orientation of signal-anchor proteins in the ER membrane seemed to be dictated to a large extent by the charge distribution in the residues that flank either side of the TM domain, with a net internal positive charge favoring an Nlumenal/ Ccytoplasmic topology. Transport of the yeast Vps10p along the prevacuolar endosome-like/vacuolar pathway requires clathrin and the adaptors Gga1p and Gga2p. Deletion of both genes impairs proteolytic processing of the inactive precursors of the vacuolar hydrolase CPY. Interestingly, Vps10p does not have the canonical DXXLL signals that are involved in the recognition by the Ggas, with the possibility that the yeast Ggas might recognize a different sorting motif. Moreover, while Vps10p contains two aromatic-based signals, YSSL and FYVF, in its cytoplasmic tail, the mutation of individual AP1 subunits or deletion of the whole complex in yeast results in no measurable protein-trafficking phenotype. We showed that the cytoplasmic YQII motif of GlVps is essential for the proper localization and stability of this receptor. Because this lysosomal motif was critical for GlVps-m1 interaction, we presumed that the AP1 complex together with clathrin might be participating in the trafficking of the receptor, similar to the transport of the lysosomal membrane protein ESCP. YQII-deleted GlVps and GlVps from m1 depleted trophozoites were proteolytically ARN509 site processed, suggesting that either the absence of the sorting motif or the lack of the adaptor counterpart this receptor might be degraded in the PVs or in the cytoplasm by proteasome. I