Please use this identifier to cite or link to this item: https://doi.org/10.1371/journal.pone.0041084
Title: Structure-function analysis of Rgs1 in magnaporthe oryzae: Role of dep domains in subcellular targeting
Authors: Ramanujam R.
Yishi X.
Liu H.
Naqvi N.I. 
Keywords: guanine nucleotide binding protein
monoclonal antibody
RGS protein
RGS1 protein
unclassified drug
fungal protein
RGS protein
amino terminal sequence
article
asexual spore
barley
carboxy terminal sequence
cell vacuole
cellular distribution
conidium
cytosol
down regulation
fungal virulence
fungus growth
fungus hyphae
hydrophobicity
immunoblotting
in vivo study
Magnaporthe
Magnaporthe grisea
Magnaporthe oryzae
microscopy
nonhuman
nucleotide sequence
pathogenesis
protein cleavage
protein degradation
protein domain
protein expression
protein function
protein motif
protein processing
protein targeting
rice
sequence alignment
signal transduction
structure activity relation
Western blotting
wettability
amino acid sequence
chemistry
Magnaporthe
metabolism
molecular genetics
sequence homology
structure activity relation
Fungi
Magnaporthe
Magnaporthe grisea
Magnaporthe oryzae
Amino Acid Sequence
Fungal Proteins
Magnaporthe
Molecular Sequence Data
RGS Proteins
Sequence Homology, Amino Acid
Structure-Activity Relationship
Issue Date: 2012
Citation: Ramanujam R., Yishi X., Liu H., Naqvi N.I. (2012). Structure-function analysis of Rgs1 in magnaporthe oryzae: Role of dep domains in subcellular targeting. PLoS ONE 7 (7) : e41084. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0041084
Rights: Attribution 4.0 International
Abstract: Background: Rgs1, a prototypical Regulator of G protein Signaling, negatively modulates the cyclic AMP pathway thereby influencing various aspects of asexual development and pathogenesis in the rice-blast fungus Magnaporthe oryzae. Rgs1 possesses tandem DEP motifs (termed DEP-A and DEP-B; for Dishevelled, Egl-10, Pleckstrin) at the N-terminus, and a G?-GTP interacting RGS catalytic core domain at the C-terminus. In this study, we focused on gaining further insights into the mechanisms of Rgs1 regulation and subcellular localization by characterizing the role(s) of the individual domains and the full-length protein during asexual development and pathogenesis in Magnaporthe. Methodology/Principal Findings: Utilizing western blot analysis and specific antisera against the N- and C-terminal halves of Rgs1, we identify and report the in vivo endoproteolytic processing/cleavage of full-length Rgs1 that yields an N-terminal DEP and a RGS core domain. Independent expression of the resultant DEP-DEP half (N-Rgs1) or RGS core (C-Rgs1) fragments, failed to complement the rgs1? defects in colony morphology, aerial hyphal growth, surface hydrophobicity, conidiation, appressorium formation and infection. Interestingly, the full-length Rgs1-mCherry, as well as the tagged N-terminal DEP domains (individually or in conjunction) localized to distinct punctate vesicular structures in the cytosol, while the catalytic RGS core motif was predominantly vacuolar. Conclusions/Significance: Based on our data from sequence alignments, immuno-blot and microscopic analysis, we propose that the post-translational proteolytic processing of Rgs1 and the vacuolar sequestration of the catalytic RGS domain represents an important means of down regulating Rgs1 function and thus forming an additional and alternative means of regulating G protein signaling in Magnaporthe. We further hypothesize the prevalence of analogous mechanisms functioning in other filamentous fungi. Furthermore, we conclusively assign a specific vesicular/membrane targeting function for the N-terminal DEP domains of Rgs1 in the rice-blast fungus. © 2012 Ramanujam et al.
Source Title: PLoS ONE
URI: https://scholarbank.nus.edu.sg/handle/10635/161971
ISSN: 19326203
DOI: 10.1371/journal.pone.0041084
Rights: Attribution 4.0 International
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