Marco Trujillo

Marco Trujillo

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68
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Publications (68)
Article
In contrast to other eukaryotic model organisms, the closely related ubiquitin (Ub)-conjugating enzymes UBC35 and UBC36 are the main sources of K63-linked Ub chains in Arabidopsis.1 Although K63-linked chains have been associated with the regulation of vesicle trafficking, definitive proof for their role in endocytosis was missing. We show that the...
Article
Full-text available
Biotrophic plant pathogens secrete effector proteins to manipulate the host physiology. Effectors suppress defenses and induce an environment favorable to disease development. Sequence-based prediction of effector function is impeded by their rapid evolution rate. In the maize pathogen Ustilago maydis, effector-coding genes frequently organize in c...
Article
During the immune response, activation of the secretory pathway is key to mounting an effective response, while gauging its output is important to maintain cellular homeostasis. The Exo70 subunit of the exocyst functions as a spatiotemporal regulator by mediating numerous interactions with proteins and lipids. However, a molecular understanding of...
Preprint
Full-text available
Biotrophic plant pathogens secrete effector proteins to manipulate the host physiology. Effectors suppress defenses and induce an environment favorable to disease development. Sequence-based prediction of effector function is difficulted by their rapid evolution rate. In the maize pathogen Ustilago maydis , effector-coding genes frequently organize...
Article
The exocyst is an evolutionary conserved complex that mediates tethering of post-Golgi vesicles derived from the conventional secretory pathway to the plasma membrane (PM), before SNARE-mediated fusion. Through its tethering function, connecting secretory vesicles to the PM, it mediates spatiotemporal regulation of exocytosis. As an integral elemen...
Preprint
The exocyst is a conserved hetero-octameric complex that mediates early tethering of post-Golgi vesicles during exocytosis. Its Exo70 subunit functions as a spatiotemporal regulator by mediating numerous interactions with proteins and lipids. However, a molecular understanding of the exocyst functions remains challenging. Exo70B2 localized to dynam...
Article
Full-text available
Ubiquitination is a prevalent post-translational modification involved in all aspects of cell physiology. It is mediated by an enzymatic cascade and the E2 ubiquitin-conjugating enzymes (UBCs) lie at its heart. Even though E3 ubiquitin ligases determine the specificity of the reaction, E2s catalyse the attachment of ubiquitin and have emerged as ke...
Article
Full-text available
Background: The large and highly repetitive genomes of the cultivated species Hordeum vulgare (barley), Triticum aestivum (wheat), and Secale cereale (rye) belonging to the Triticeae tribe of grasses appear to be particularly rich in gene-like sequences including partial duplicates. Most of them have been classified as putative pseudogenes. In thi...
Article
Full-text available
The role of reactive oxygen intermediate (ROI) accumulation in resistance and susceptibility of plants to parasitic fungi is still little understood. We examined the spatial and temporal occurrence of different ROIs in barley after inoculation with the biotrophic fungus Blumeria graminis f.sp. hordei (Bgh, barley powdery mildew fungus). Using histo...
Article
Plant U-box type E3 ubiquitin ligases (PUBs) are well known for their functions in a variety of stress responses, including immune responses and the adaptation to abiotic stresses. First linked to pollen self-incompatibility, their repertoire of roles has grown to encompass also the regulation of developmental processes. Notably, new studies provid...
Article
Ubiquitination is mediated by an enzymatic cascade that results in the modification of substrate proteins, redefining their fate. This post‐translational modification is involved in most cellular processes, yet its analysis faces manifold obstacles due to its complex and ubiquitous nature. Reconstitution of the ubiquitination cascade in bacterial s...
Article
Full-text available
The induction of rapid cell death is an effective strategy for plants to restrict biotrophic and hemi-biotrophic pathogens at the infection site. However, activation of cell death comes at a high cost, as dead cells will no longer be available for defense responses nor general metabolic processes. In addition, necrotrophic pathogens that thrive on...
Data
The GYF domain proteins. a, Aligned amino acid sequences of GYF domains from diverse eukaryotic organisms. Key residues for GYF domains are delineated as white text on a black or gray background. The conserved Ile residue of Ile-type GYF domains are indicated in bold text. At, Os, Smo, Phpat, Cre, Kfl, Hs, and Sc stand for following species: Arabid...
Data
Stomatal density and stomatal closure response. a, Flg22-induced stomatal closure in the psig1-1 mutant. Stomatal apertures were measured following treatment with 5 μM flg22 or mock (dH2O) for 60 min. Data are shown as the mean ± SE (n = 24). Statistical groups were determined using the Tukey HSD test. Statistically significant differences are indi...
Data
Pto syringe infiltration test in the psig1 mutants. 5-week-old plants were syringe inoculated with 5 x 104 c.f.u. ml-1 of Pto under long day conditions (12 h light / 12 h dark), and bacterial growth was determined at 0 and 3 dpi for a and at 0, 2 and 3 dpi for b. Data are shown as the mean ± SE. Two leaves were taken from each of 3 individual plant...
Data
Pto AvrRPS4 infiltration test in the psig1 mutants. Plants were syringe inoculated with 5 x 105 c.f.u. ml-1 of Pto AvrRPS4, and bacterial growth was determined at 3 dpi. Data are shown as the mean ± SE. Statistical groups were determined using the Tukey HSD test. Statistically significant differences are indicated by different letters (p < 0.05). (...
Data
PCR primers used in this study. (PDF)
Data
The psig1 mutant phenotypes. a and b, The psig1 mutants were more susceptible to Pto. Plants were spray inoculated with 1 x 108 c.f.u. ml-1 of Pto, and bacterial growth was determined at 0 and 2 or 3 dpi. Data are shown as the mean ± SE. Statistical groups were determined using the Tukey HSD test. Statistically significant differences are indicated...
Data
The psig1 mutants displayed enhanced cell death upon infection with Pto AvrRPM1 or Pto AvrRPS4. a, c, d and f, Plants were spray inoculated with 1 x 108 c.f.u. ml-1 of Pto AvrRPM1 or Pto AvrRPS4, and dead cells were visualized by trypan blue staining 1 or 2 day after inoculation. The scale bar represents 200 μm. b, e and g, Trypan blue stained area...
Data
The psig1 eds1-2 (Col) mutant displayed enhanced cell death upon infection with Pto AvrRPM1. a, Plants were dip inoculated with 1 x 108 c.f.u. ml-1 of Pto AvrRPM1, and dead cells were visualized by trypan blue staining 1 day after inoculation. The scale bar represents 200 μm. b, Trypan blue stained area. Plants were spray inoculated with 1 x 108 c....
Data
PSIG1 is associated with DCP1 and SMG7. The images show the GFP signal in green and the mCherry or TagRFP signal in red. The merged images indicate the overlay of two signals in yellow. a, Subcellular localization was analyzed at 4 days after inoculation in agroinfiltrated Nicotiana benthamiana. b, Subcellular localization was analyzed at 3 days af...
Data
PSIG1 gene expression in the WT Col-8 plants. a, PSIG1 gene expression upon in leaves of soil grown plants. Six-week-old plants were syringe infiltrated with 1 x 106 c.f.u. ml-1 of Pto. b, PSIG1 gene expression in 10-day-old liquid culture grown seedlings. The data are shown as mean ± SE. No significant differences were observed between non-treated...
Data
Phenotypes of complemented lines. a, Photograph of 7 week-old plants grown under short day conditions. b, PSIG1 gene expression in 6-week-old plants. Data are shown as the mean ± SE. Statistical groups were determined using the Tukey HSD test. Statistically significant differences are indicated by different letters (p < 0.05). c, Flg22-induced ROS...
Data
Cell death induction in the bak1-4 mutant. a, Plants were inoculated with Hpa Noco2 spores, and dead cells on the true leaves were visualized by trypan blue staining 6 days after inoculation. The scale bar represents 200 μm. b, Plants were spray inoculated with 1 x 108 c.f.u. ml-1 of Pto AvrRPS4, and dead cells were visualized by trypan blue staini...
Data
The PPGF sequence of SMG7 is required for interaction with PSIG1 in vitro. HisMBP-SMG7 or HisMBP-SMG7G933A was incubated with GST, GST-PSIG11-606, or GST-PSIG11-606 Y575A, and the conjugates were pulled down with Glutathione-Sepharose beads. HisMBP-SMG7 and GST-PSIG1 were detected by immunoblotting using anti-His antibody or anti-GST antibody. (TIF...
Data
The smg7-4 mutant allele displayed enhanced cell death upon infection with Pto AvrRPM1. a, Plants were spray inoculated with 1 x 108 c.f.u. ml-1 of Pto AvrRPM1, and dead cells were visualized by trypan blue staining 1 day after inoculation. The scale bar represents 200 μm. b and c, Trypan blue stained area. Plants were spray inoculated with 1 x 108...
Data
MSMS spectrum for the phosphopeptide 'DIQGSDNAIPLpSPQWLLSKPGENK'. (PDF)
Data
Protein Identity of PSIG homologs. (TIF)
Data
R script of statistical analysis of quantitative PCR data using R. (PDF)
Data
R script of Tukey HSD test and visualization of box plot using ggplot2. (PDF)
Data
The psig1 T-DNA insertion mutants. a, The psig1 mutant alleles display a slight dwarf phenotype. Photograph of 6-week-old plants grown under short day conditions. b, Genomic structure of the PSIG1 gene and the position of the T-DNA insert. Exons are indicated as black boxes. The T-DNA insertion sites are indicated by grey triangles. Black arrowhead...
Data
Flg22 responses in the psig1 mutants. a, Flg22-induced ROS production in the psig1 mutants. Data are shown as the mean ± SE. b, Flg22-induced MAPK activation in the psig1 mutants. c, Flg22-induced callose deposition in the psig1 mutants. The scale bar represents 200 μm. d, Callose deposition was quantified with Image J software. Data are shown as t...
Data
PR1 gene expression in the psig1 mutants. a, PR1 gene expression in 10-day-old liquid culture grown seedlings. Expression data for PR1 are shown as the mean ± SE. Statistical groups were determined using the Tukey HSD test. Statistically significant differences are indicated by different letters (p < 0.05). b, PR1 gene expression in leaves of soil...
Data
Pto AvrRPM1 induced cell death was suppressed by pre-treatment of flg22 in psig1-1. Trypan blue stained area. Pretreated plants either with 1 μM flg22 or dH2O (mock) were spray inoculated with 1 x 108 c.f.u. ml-1 of Pto AvrRPM1, and dead cells were visualized by trypan blue staining 1 day after inoculation. The stained area was measured using an im...
Data
Plant SMG7 homologs have the PPGF sequence in the non-conserved C-terminal region. a, Phylogenetic tree and schematic structures of SMG7 from diverse eukaryotic organisms. Numbers on the phylogenetic tree indicate the bootstrap values. Dark blue boxes indicate the EST1 domain. Yellow, green and blue boxes indicate the Est1 DNA/RNA binding domain, t...
Data
Protein similarity of PSIG homologs. (TIF)
Data
Selected phosphoproteomics data. (XLSX)
Article
Full-text available
In plant-pathogen interactions, components of the plant ubiquitination machinery are preferred targets of pathogen-encoded effectors suppressing defense responses or co-opting host cellular functions for accommodation. Here, we employed transient and stable gene silencing- and over-expression systems in Hordeum vulgare (barley) to study the functio...
Article
Full-text available
Auxin is a small molecule morphogen that bridges SCFTIR1/AFB-AUX/IAA co-receptor interactions leading to ubiquitylation and proteasome-dependent degradation of AUX/IAA transcriptional repressors. Here, we systematically dissect auxin sensing by SCFTIR1-IAA6 and SCFTIR1-IAA19 co-receptor complexes, and assess IAA6/IAA19 ubiquitylation in vitro and I...
Data
Supplementary Figures, Supplementary Tables, Supplementary Note and Supplementary References
Data
Sequence alignment of 29 A. thaliana AUX/IAA proteins generated for the phylogenetic tree in Supplementary Fig. 1.
Data
Multiple sequence alignments of AUX/IAAs from 81 A. thaliana accessions and A. halleri, A. lyrata, and C. rubella
Article
Full-text available
Crosstalk between post-translational modifications such as ubiquitination and phosphorylation play key roles in controlling the duration and intensity of signalling events to ensure cellular homeostasis. However, the molecular mechanisms underlying the regulation of negative feedback loops remain poorly understood. Here we uncover a pathway in Arab...
Chapter
Ubiquitination is a central posttranslational modification that impinges on the fate of proteins. While attachment of K48-linked chains onto soluble proteins marks them for proteolysis via the 26S proteasome, mono-ubiquitination or K63-linked chains result in the endocytosis and sorting through the endomembrane system of integral membrane proteins,...
Chapter
The rapid production of reactive oxygen species (ROS) in response to biotic and abiotic cues is a conserved hallmark of plant responses. The detection and quantification of ROS generation during immune responses is an excellent readout to analyze signaling triggered by the perception of pathogens. The assay described here is easy to employ and vers...
Article
Full-text available
Targeted proteomics has become increasingly popular recently because of its ability to precisely quantify selected proteins in complex cellular backgrounds. We demonstrate the utility of an LTQ-Orbitrap Velos Pro mass spectrometer in targeted parallel reaction monitoring (PRM) despite its unconventional dual ion trap configuration. We evaluated abs...
Article
Plant cells dynamically change their architecture and molecular composition following encounters with beneficial or parasitic microbes, a process referred to as host cell reprogramming. Cell-autonomous defense reactions are typically polarized to the plant cell periphery underneath microbial contact sites, including de novo cell wall biosynthesis....
Article
Full-text available
Components of the vesicle trafficking machinery are central to the immune response in plants. The role of vesicle trafficking during pre-invasive penetration resistance has been well documented. However, emerging evidence also implicates vesicle trafficking in early immune signaling. Here we report that Exo70B1, a subunit of the exocyst complex whi...
Article
Full-text available
Plant pathogens are perceived by pattern recognition receptors, which are activated upon binding to pathogen-associated molecular patterns (PAMPs). Ubiquitination and vesicle trafficking have been linked to the regulation of immune signaling. However, little information exists about components of vesicle trafficking involved in immune signaling and...
Article
Full-text available
From pathogen perception and the activation of signal transduction cascades to the deployment of defense responses, protein ubiquitination plays a key role in the modulation of plant immunity. Ubiquitination is mediated by three enzymes, of which the E3 ubiquitin ligases, the substrate determinants, have been the major focus of attention. Accumulat...
Article
Full-text available
Piriformospora indica is a root-colonizing basidiomycete that confers a wide range of beneficial traits to its host. The fungus shows a biotrophic growth phase in Arabidopsis (Arabidopsis thaliana) roots followed by a cell death-associated colonization phase, a colonization strategy that, to our knowledge, has not yet been reported for this plant....
Article
Plant immune responses require the coordination of a myriad of processes that are triggered upon perception of invading pathogens. Ubiquitin, the ubiquitination system (UBS) and the 26S proteasome are key for the regulation of processes such as the oxidative burst, hormone signaling, gene induction, and programmed cell death. E3 ligases, the specif...
Article
The first line of active defense in plants is triggered by invariant microbial epitopes known as pathogen-associated molecular patterns (PAMPs). Perception of PAMPs by receptors activates a plethora of reactions ending in PAMP-triggered immunity (PTI), which contributes to broad-spectrum resistance [1 • Jones J.D. • Dangl J.L. The plant immune sys...
Article
Reactive oxygen intermediates (ROI) are closely related to defence reactions of plants against pathogens. A prominent role in the production of ROI has been attributed to the plant respiratory burst oxidase homologues (RBOH) of the human phagocyte GP91(phox). A barley RBOH, which encodes a putative superoxide (O2·−) producing NADPH oxidase, is desc...
Article
SUMMARY Non-host resistance of barley to Blumeria graminis f.sp. tritici (Bgt), an inappropriate forma specialis of the grass powdery mildew fungus, is associated with formation of cell wall appositions (papillae) at sites of attempted fungal penetration and a hypersensitive cell death reaction (HR) of single attacked cells. Penetration resistance...
Article
Full-text available
Nonhost resistance of cereals to inappropriate formae speciales of Blumeria graminis is little understood. However, on the microscopic level, nonhost defense to B. graminis is reminiscent of host defense preventing fungal development by penetration resistance and the hypersensitive cell death response (HR). We analyzed histochemically the accumulat...
Article
Full-text available
We analysed pathogenesis-related expression of genes, that are assumed to be involved in ubiquitous plant defence mechanisms like the oxidative burst, the hypersensitive cell death reaction (HR) and formation of localized cell wall appositions (papillae). We carried out comparative northern blot and RT-PCR studies with near-isogenic barley (Hordeum...
Article
Full-text available
The pathogenesis-related accumulation of superoxide radical anions (O2*-) and hydrogen peroxide (H2O2) was comparatively analyzed in a barley line (Hordeum vulgare L. cv Sultan-5) carrying the powdery mildew (Blumeria graminis f.sp. hordei, Speer, Bgh) resistance gene Mla12, and in susceptible mutants defective in Mla12 or in genes "required for Ml...
Article
Abstract Race nonspecific resistance of barley against the barley powdery mildew fungus (Blumeria Graminis f.sp. Hordei, Speer, Bgh) is mediated by recessive mlo alleles and is controlled by at least two additional genes 'required for ml o-specified disease resistance' (Ror1 and Ror2). The pathogenesis-related accumulation of hydrogen peroxide (H(2...
Conference Paper
The U-box E3 ligase is a key regulatory component of plant ubiquitination machinery affecting almost all aspects of plants growth and development. Because of its central role in degradation and modification of target proteins, E3 ligases are also preferred targets of pathogen-encoded effectors for manipulation of host’s defense responses. Plants on...

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