A membrane-associated movement protein of Pelargonium flower break virus shows RNA-binding activity and contains a biologically relevant leucine zipper-like motif

[EN] Two small viral proteins (DGBp1 and DGBp2) have been proposed to act in a concerted manner to aid intra- and intercellular trafficking of carmoviruses though the distribution of functions and mode of action of each protein partner are not yet clear. Here we have confirmed the requirement of the DGBps of Pelargonium flower break virus (PFBV), p7 and p12, for pathogen movement Studies focused on p12 have shown that it associates to cellular membranes, which is in accordance to its hydrophobic profile and to that reported for several homologs. However, peculiarities that distinguish p12 from other DGBp52 have been found. Firstly, it contains a leucine zipper-like motif which is essential for virus infectivity in plants. Secondly, it has an unusually long and basic N-terminal region that confers RNA binding activity. The results suggest that PFBV p12 may differ mechanistically from related proteins and possible roles of PFBV DGBps are discussed. (C) 2011 Elsevier Inc. All rights reserved.We gratefully thank Dr. Vicente Pallas for critical reading of the manuscript and Dolores Arocas and Isabella Avellaneda for their technical assistance. This research was supported by grant BFU2006-11230 and BFU2009-11699 from the Ministerio Ciencia e Innovacion (MICINN, Spain) and by grants ACOM/2006/210 and ACOMP/2009/040 (Generalitat Valenciana, GV) to C. H. S. M.-T. was the recipient of a predoctoral fellowship from GV and of a predoctoral contract from MICINN.Martínez Turiño, S.; Hernandez Fort, C. (2011). A membrane-associated movement protein of Pelargonium flower break virus shows RNA-binding activity and contains a biologically relevant leucine zipper-like motif. Virology. 413(2):310-319. https://doi.org/10.1016/j.virol.2011.03.001S310319413

Yeast Two-Hybrid, a Powerful Tool for Systems Biology

A key property of complex biological systems is the presence of interaction networks formed by its different components, primarily proteins. These are crucial for all levels of cellular function, including architecture, metabolism and signalling, as well as the availability of cellular energy. Very stable, but also rather transient and dynamic protein-protein interactions generate new system properties at the level of multiprotein complexes, cellular compartments or the entire cell. Thus, interactomics is expected to largely contribute to emerging fields like systems biology or systems bioenergetics. The more recent technological development of high-throughput methods for interactomics research will dramatically increase our knowledge of protein interaction networks. The two most frequently used methods are yeast two-hybrid (Y2H) screening, a well established genetic in vivo approach, and affinity purification of complexes followed by mass spectrometry analysis, an emerging biochemical in vitro technique. So far, a majority of published interactions have been detected using an Y2H screen. However, with the massive application of this method, also some limitations have become apparent. This review provides an overview on available yeast two-hybrid methods, in particular focusing on more recent approaches. These allow detection of protein interactions in their native environment, as e.g. in the cytosol or bound to a membrane, by using cytosolic signalling cascades or split protein constructs. Strengths and weaknesses of these genetic methods are discussed and some guidelines for verification of detected protein-protein interactions are provided

A conserved major facilitator superfamily member orchestrates a subset of O-glycosylation to aid macrophage tissue invasion

Aberrant display of the truncated core1 O-glycan T-antigen is a common feature of human cancer cells that correlates with metastasis. Here we show that T-antigen in Drosophila melanogaster macrophages is involved in their developmentally programmed tissue invasion. Higher macrophage T-antigen levels require an atypical major facilitator superfamily (MFS) member that we named Minerva which enables macrophage dissemination and invasion. We characterize for the first time the T and Tn glycoform O-glycoproteome of the Drosophila melanogaster embryo, and determine that Minerva increases the presence of T-antigen on proteins in pathways previously linked to cancer, most strongly on the sulfhydryl oxidase Qsox1 which we show is required for macrophage tissue entry. Minerva’s vertebrate ortholog, MFSD1, rescues the minerva mutant’s migration and T-antigen glycosylation defects. We thus identify a key conserved regulator that orchestrates O-glycosylation on a protein subset to activate a program governing migration steps important for both development and cancer metastasis

The Marble Faun

Undergraduate student, year of graduation2018 . Major: Painting. Class: Drawing I. Faculty: James Stanley.https://digitalcommons.risd.edu/bookcontest2nd2016/1249/thumbnail.jp

The Marble Faun

Undergraduate student, year of graduation2018 . Major: Painting. Class: Drawing I. Faculty: James Stanley.https://digitalcommons.risd.edu/bookcontest2nd2016/1246/thumbnail.jp

The Marble Faun

Undergraduate student, year of graduation2018 . Major: Painting. Class: Drawing I. Faculty: James Stanley.https://digitalcommons.risd.edu/bookcontest2nd2016/1247/thumbnail.jp

The Marble Faun

Undergraduate student, year of graduation2018 . Major: Painting. Class: Drawing I. Faculty: James Stanley.https://digitalcommons.risd.edu/bookcontest2nd2016/1245/thumbnail.jp