A reduced VWA domain-containing proteasomal ubiquitin receptor of Giardia lamblia localizes to the flagellar pore regions in microtubule-dependent manner

BACKGROUND: Giardia lamblia switches its lifecycle between trophozoite and cyst forms and the proteasome plays a pivotal role in this switching event. Compared to most model eukaryotes, the proteasome of this parasite has already been documented to have certain variations. This study was undertaken to characterize the ubiquitin receptor, GlRpn10, of the 19S regulatory particle of the Giardia proteasome and determine its cellular localization in trophozoites, encysting trophozoites and cysts. METHOD: Sequence alignment and domain architecture analyses were performed to characterize GlRpn10. In vitro ubiquitin binding assay, functional complementation and biochemical studies verified the protein’s ability to function as ubiquitin receptor in the context of the yeast proteasome. Immunofluorescence localization was performed with antibody against GlRpn10 to determine its distribution in trophozoites, encysting trophozoites and cysts. Real-time PCR and Western blotting were performed to monitor the expression pattern of GlRpn10 during encystation. RESULT: GlRpn10 contained a functional ubiquitin interacting motif, which was capable of binding to ubiquitin. Although it contained a truncated VWA domain, it was still capable of partially complementing the function of the yeast Rpn10 orthologue. Apart from localizing to the nucleus and cytosol, GlRpn10 was also present at flagellar pores of trophozoites and this localization was microtubule-dependent. Although there was no change in the cellular levels of GlRpn10 during encystation, its selective distribution at the flagellar pores was absent. CONCLUSION: GlRpn10 contains a noncanonical VWA domain that is partially functional in yeast. Besides the expected nuclear and cytosolic distribution, the protein displays microtubule-dependent flagellar pore localization in trophozoites. While the protein remained in the nucleus and cytosol in encysting trophozoites, it could no longer be detected at the flagellar pores. This absence at the flagellar pore regions in encysting trophozoites is likely to involve redistribution of the protein, rather than decreased gene expression or selective protein degradation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-015-0737-1) contains supplementary material, which is available to authorized users

A new typothoracine aetosaur (Archosauria, Pseudosuchia) from the Upper Triassic of India with insights on biostratigraphy, diversification, and paleobiogeography

A new typothoracine aetosaur is described based on multiple isolated and articulated left paramedian and lateral osteoderms recovered from the Upper Triassic lower Dharmaram Formation of India. The partial carapace of the new taxon is reconstructed as strongly discoidal based on the curvature of the paramedian osteoderms with the widest one positioned dorsal to the mid-dorsal trunk vertebra. Asymmetric lateral osteoderms with acute flexion are considered as precaudals with the angle of flexion decreasing posteriorly. Phylogenetic analysis recovered the new taxon as deeply nested within the clades Typothoracinae and Paratypothoracini, and a sister taxon to Kocurypelta silvestris. The autapomorphic characters involving paramedian osteoderms dorsal to the trunk vertebrae include dorsal surface ornamented by large, irregular pits surrounding the dorsal eminence and radiating ridges in other areas, straight anterior margin of the anteromedial corner of the anterior bar in dorsal view, and raised or ridged and ornamented posteromedial corner. The current study highlights the significance of the new aetosaur, and the age of the lower Dharamaram Formation is modified here as mid-Norian to Rhaetian based on global correlation with other coeval horizons. The recovery of this taxon marks the first record of Paratypothoracini from high paleolatitudes of the Gondwanan region. The study corroborates the earlier findings of a strong Laurasian faunal influx in India during the Late Triassic suggesting possible land bridges and/or conducive environmental conditions for faunal dispersal. http://zoobank.org/urn:lsid:zoobank.org:pub:FCC37C0E-107F-4B15-9460-4E802B465865</p

Additional file 3: of Variation in the ribosome interacting loop of the Sec61α from Giardia lamblia

Tertiary structure of GlSec61α obtained by homology modeling based on 2WWB (a & b) and 3J7Q (c & d), followed by molecular dynamic simulation for 30 ns, with (b & d) or without (a & c) docked RNA. Other details are same as those described in Fig. 1b. The corresponding RMSD graphs are shown below each structure. The templates used for modeling (2WWB and 3J7Q) had several other protein/peptide chains and RNA fragments, all of which exerted a constraint on the protein conformation. Thus, although the homology modeled structures resembled that conformation of the template, the above-mentioned constraints were absent in our simulations as only a fragment of the RNA was used for docking. As a result, during simulation, the structures relaxed in the first few nanoseconds and this resulted in a rise of the RMSD before subsequent stabilization. Hence observed increase of RMSD does not reflect any destabilization. (PPTX 1691 kb

La Petite presse : journal quotidien... / [rédacteur en chef : Balathier Bragelonne]

17 novembre 18791879/11/17 (A13,N4941)