Journal: Molecules

Article Title: Metabolomic Insights into the Antimicrobial Effects of Metschnikowia Yeast on Phytopathogens

doi: 10.3390/molecules30153268

Figure Lengend Snippet: Identification via LARAPPI/CI-MSI analysis of chemical compounds synthesized by Metschnikowia pulcherrima yeast strains D2, D4, and TK1 in control samples and during co-cultivation with molds ( Botrytis cinerea , Rhizoctonia solani , Alternaria alternata , Monilia laxa ).

Article Snippet: Alternaria tenuissima DSM 63360, Colletotrichum coccodes DSM 62126, Fusarium sambucinum DSM 62397, Phoma exigua DSM 62040, and Rhizoctonia solani DSM 22843 were obtained from Collection of Pure Cultures of the Leibniz Institute DSMZ (Germany).

Techniques: Synthesized, Control

Journal: Molecules

Article Title: Metabolomic Insights into the Antimicrobial Effects of Metschnikowia Yeast on Phytopathogens

doi: 10.3390/molecules30153268

Figure Lengend Snippet: Statistical analysis of the entire MS data set. ( A ) Principal Component Analysis (PCA) score plot of metabolites from cultures of M. pulcherrima yeast D2, D4, and TK1 (controls: KD2, KD4, KTK11) and co-cultures of M. pulcherrima yeast strains D2, D4, and TK1 with phytopathogenic molds (P5— Rhizoctonia solani , P9— Botrytis cinerea , P10— Monilia laxa , P11— Alternaria alternata ); ( B ) heatmap of metabolite abundance, distinguishing cultures of M. pulcherrima yeast strains D2, D4, and TK1, and co-cultures of M. pulcherrima yeast strains D2, D4, and TK1 with phytopathogenic molds (P5— R. solani , P9— B. cinerea , P10— M. laxa , P11— A. alternata ). Red indicates higher relative abundance; blue indicates lower relative abundance. The color bar on the right represents the class labels for samples. Analysis performed via UHPLC-QToF-UHRMS.

Article Snippet: Alternaria tenuissima DSM 63360, Colletotrichum coccodes DSM 62126, Fusarium sambucinum DSM 62397, Phoma exigua DSM 62040, and Rhizoctonia solani DSM 22843 were obtained from Collection of Pure Cultures of the Leibniz Institute DSMZ (Germany).

Techniques:

Journal: Molecules

Article Title: Synthesis and Biological Properties of Fluorescent Strigolactone Mimics Derived from 1,8-Naphthalimide

doi: 10.3390/molecules29102283

Figure Lengend Snippet: The effect of SL mimics on the growth of phytopathogens expressed as colony diameter on agar plates: ( a ) Fusarium graminearum DSM 4527; ( b ) Rhizoctonia solani DSM 22842; ( c ) Sclerotinia sclerotium DSM 1946; ( d ) Colletotrichum acutatum CBS 113008; C1 = 5 × 10 −6 M; C2 = 10 −5 M; C3 = 5 × 10 −5 M. The bars represent the standard error. Significant differences are shown by different letters at p < 0.05 ( n = 3).

Article Snippet: The fungal strains tested were represented by C. acutatum CBS 113008 (Culture collection of fungi and yeasts, Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherland), F. graminearum DSM 4527, R. solani DSM 22842, and S. sclerotium DSM 1946 (DSMZ, Braunschweig, Germany).

Techniques:

Journal: Molecules

Article Title: Synthesis and Biological Properties of Fluorescent Strigolactone Mimics Derived from 1,8-Naphthalimide

doi: 10.3390/molecules29102283

Figure Lengend Snippet: Hyphal branching of Fusarium graminearum DSM 4527 ( a ), Rhizoctonia solani DSM 22842 ( b ), Sclerotinia sclerotium DSM 1946 ( c ), and Colletotrichum acutatum CBS 113008 ( d ) exposed to GR24 at concentration C3 (5 × 10 −5 M); the arrows indicate the following: 1—primary branch; 2—2nd-order branch; 3—3rd-order branch; 4—4th-order branch; 5—5th-order branch.

Article Snippet: The fungal strains tested were represented by C. acutatum CBS 113008 (Culture collection of fungi and yeasts, Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherland), F. graminearum DSM 4527, R. solani DSM 22842, and S. sclerotium DSM 1946 (DSMZ, Braunschweig, Germany).

Techniques: Concentration Assay

Journal: Molecules

Article Title: Synthesis and Biological Properties of Fluorescent Strigolactone Mimics Derived from 1,8-Naphthalimide

doi: 10.3390/molecules29102283

Figure Lengend Snippet: The effect of GR24 and SL mimics on the number of hyphal branches of phytopathogens: ( a ) Fusarium graminearum DSM 4527; ( b ) Rhizoctonia solani DSM 22842; ( c ) Sclerotinia sclerotium DSM 1946; ( d ) Colletotrichum acutatum CBS 113008; C1 = 5×10 −6 M; C2 = 10 −5 M; C3 = 5 × 10 −5 M. The bars represent the standard error. Significant differences are shown by different letters at p < 0.05 ( n = 30).

Article Snippet: The fungal strains tested were represented by C. acutatum CBS 113008 (Culture collection of fungi and yeasts, Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherland), F. graminearum DSM 4527, R. solani DSM 22842, and S. sclerotium DSM 1946 (DSMZ, Braunschweig, Germany).

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Human C1orf27 protein interacts with α 2A -adrenergic receptor and regulates its anterograde transport

doi: 10.1016/j.jbc.2022.102021

Figure Lengend Snippet: siRNA-mediated depletion of C1orf27 impedes the ER-to-Golgi transport kinetics of nascent α 2A -AR . A , Western blot analysis of siRNA-mediated knockdown of endogenous C1orf27. B , effect of C1orf27 knockdown by siRNA on the ER–Golgi transport kinetics of α 2A -AR. HeLa cells were transfected with Str-KDEL_SBP-EGFP-α 2A -AR plasmids together with control or C1orf27 siRNA. α 2A -AR transport from the ER was induced after addition of biotin at 0 min. C , quantitative data showing the normalized Golgi/total expression of α 2A -AR over time in control or C1orf27 siRNA-transfected cells. After addition of biotin, images were captured at an interval of 1 min. The Golgi/total ratio at each time was normalized to the highest ratio after subtraction from the ratio at time 0 in individual cells. D , the half time of ER–Golgi transport of α 2A -AR in control and C1orf27 siRNA-transfected cells. The quantitative data are mean ± SE (n = 20–26 cells in 6–10 individual experiments). ∗∗∗ p < 0.001 versus control siRNA. Scale bars, 10 μm. α2A-AR, α2A-adrenergic receptor; ER, endoplasmic reticulum.

Article Snippet: Antibodies against C1orf27 were purchased from Proteintech.

Techniques: Western Blot, Knockdown, Transfection, Control, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Human C1orf27 protein interacts with α 2A -adrenergic receptor and regulates its anterograde transport

doi: 10.1016/j.jbc.2022.102021

Figure Lengend Snippet: C1orf27 depletion inhibits the transport of α 2A -AR from the ER to the Golgi in different cell types. A , effect of C1orf27 knockdown by siRNA on α 2A -AR export from the ER to the Golgi in RUSH assays in fixed cells. HeLa or HEK293 cells were transfected with Str-KDEL_SBP-EGFP-α 2A -AR plasmids together with control or C1orf27 siRNA and fixed at 15 and 30 min after addition of biotin. B , quantitative data shown in A. The quantitative data are the Golgi/total ratio and expressed as mean ± SE (n = 15–24 cells in 3–4 experiments). ∗∗∗ p < 0.001 versus control siRNA. Scale bars, 10 μm. α2A-AR, α2A-adrenergic receptor; ER, endoplasmic reticulum; RUSH, retention using the selective hooks.

Article Snippet: Antibodies against C1orf27 were purchased from Proteintech.

Techniques: Knockdown, Transfection, Control

Journal: The Journal of Biological Chemistry

Article Title: Human C1orf27 protein interacts with α 2A -adrenergic receptor and regulates its anterograde transport

doi: 10.1016/j.jbc.2022.102021

Figure Lengend Snippet: C1orf27 depletion attenuates the surface transport and signaling of α 2A -AR . A , schematic diagram showing modified BRET assays to measure the cell surface transport of nascent α 2A -AR after synthesis in the ER and induction with biotin using the RUSH system. B , the surface transport of α 2A -AR as measured in live cell RUSH-based BRET assays. HEK293 cells were transfected with Str-KDEL_SBP-α 2A -AR-Rluc8 and Venus-kRas for 20 h and then induced with biotin. The surface expression of α 2A -AR was measured by BRET assays. C , ERK1/2 activation by Str-KDEL_SBP-α 2A -AR-Rluc8. HEK293 cells transfected with Str-KDEL_SBP-α 2A -AR-Rluc8 or control plasmids were treated with biotin for 2 h and then stimulated with UK14304 at 1 μM for 5 min. D , quantitative data shown in C. E , inhibition of the surface transport of α 2A -AR by C1orf27 siRNA as measured in live cell RUSH-based BRET assays. HEK293 cells were transfected with Str-KDEL_SBP-α 2A -AR-Rluc8 and Venus-kRas together with control or C1orf27 siRNA. After incubation with biotin for 2 h, the surface expression of α 2A -AR was measured by BRET assays. F , inhibition of the surface expression of α 2A -AR by C1orf27 siRNA as measured in ELISA. G , effect of C1orf27 knockdown on α 2A -AR–mediated ERK1/2 activation. HEK293 cells were transfected with α 2A -AR together with control or C1orf27 siRNA and stimulated with UK14304 at 1 μM for 5 min. H , quantitative data shown in G. The quantitative data are mean ± SE (n = 3). ∗ p < 0.05, ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus control. α2A-AR, α2A-adrenergic receptor; BRET, bioluminescence resonance energy transfer; ER, endoplasmic reticulum; ERK1/2, extracellular signal–regulated kinase 1 and 2; PM, plasma membrane; RUSH, retention using the selective hooks.

Article Snippet: Antibodies against C1orf27 were purchased from Proteintech.

Techniques: Modification, Transfection, Expressing, Activation Assay, Control, Inhibition, Incubation, Enzyme-linked Immunosorbent Assay, Knockdown, Bioluminescence Resonance Energy Transfer, Clinical Proteomics, Membrane

Journal: The Journal of Biological Chemistry

Article Title: Human C1orf27 protein interacts with α 2A -adrenergic receptor and regulates its anterograde transport

doi: 10.1016/j.jbc.2022.102021

Figure Lengend Snippet: CRISPR-Cas9–mediated C1orf27 KO suppresses α 2A -AR export from the ER to the Golgi and the cell surface . A and B , inhibition of ER–Golgi transport of α 2A -AR in HeLa ( A ) and SHSY5Y cells ( B ) as measured in RUSH assays. The cells were transfected with Str-KDEL_SBP-mCherry-α 2A -AR together with control ( upper panel ) or C1orf27 KO plasmids ( lower panel ) and fixed at 15 or 30 min after addition of biotin. Inserts show GFP expression. Arrows indicate cells without transfection of C1orf27 KO plasmids in which α 2A -AR was exported to the Golgi. C , quantitative data shown in A and B. D , C1orf27 KO abolishes the surface transport of stably expressed α 2A -AR. HEK293 cells stably expressing HA-α 2A -AR were transfected with control or C1orf27 KO plasmids and stained with HA antibodies in nonpermeabilized cells. E , intracellular accumulation of α 2A -AR in cells expressing C1orf27 KO plasmids in HEK293 cells. The cells were transfected with α 2A -AR-RFP together control ( upper panel ) or C1orf27 KO plasmids ( lower panel ) carrying GFP. Arrows indicate cells without transfection with control plasmids ( upper panel ) or C1orf27 KO plasmids ( lower panel ) in which α 2A -AR was expressed at the surface. The quantitative data shown are the Golgi/total expression ratio and expressed as mean ± SE (n = 16–20 cells in three separate experiments). ∗∗∗ p < 0.001 versus control. Scale bars, 10 μm; Scale bars for inserts, 20 μm. α2A-AR, α2A-adrenergic receptor; ER, endoplasmic reticulum; GFP, green fluorescent protein; HA, hemagglutinin; KO, knockout; RFP, red fluorescent protein; RUSH, retention using the selective hooks.

Article Snippet: Antibodies against C1orf27 were purchased from Proteintech.

Techniques: CRISPR, Inhibition, Transfection, Control, Expressing, Stable Transfection, Staining, Knock-Out

Journal: The Journal of Biological Chemistry

Article Title: Human C1orf27 protein interacts with α 2A -adrenergic receptor and regulates its anterograde transport

doi: 10.1016/j.jbc.2022.102021

Figure Lengend Snippet: C1orf27 depletion inhibits the export of β 2 -AR and D2R, but not EGFR, from the ER through the Golgi to the cell surface . A – C , effect of C1orf27 knockdown by siRNA on the ER–Golgi transport of β 2 -AR ( A ), D2R ( B ), and EGFR ( C ) in RUSH assays in fixed cells. HeLa cells were transfected with individual receptor plasmids together with control or C1orf27 siRNA and fixed at 15 and 30 min after addition of biotin. Scale bars, 10 μm. D , quantitative data shown in A–C . E , effect of C1orf27 siRNA on the surface expression of α 2A -AR, β 2 -AR, D2R, and EGFR as measured in BRET assays. The quantitative data are expressed as mean ± SE (n = 17–26 cells in three experiments in D and n = 3 in E ). ∗ p < 0.05 and ∗∗∗ p < 0.001 versus control siRNA. α2A-AR, α2A-adrenergic receptor; β2-AR, β2-adrenergic receptor; BRET, bioluminescence resonance energy transfer; D2R, dopamine D2 receptor; EGF, epidermal growth factor; EGFR, EGF receptor; RUSH, retention using the selective hooks.

Article Snippet: Antibodies against C1orf27 were purchased from Proteintech.

Techniques: Knockdown, Transfection, Control, Expressing, Bioluminescence Resonance Energy Transfer

Journal: The Journal of Biological Chemistry

Article Title: Human C1orf27 protein interacts with α 2A -adrenergic receptor and regulates its anterograde transport

doi: 10.1016/j.jbc.2022.102021

Figure Lengend Snippet: C1orf27 interaction with α 2A -AR and identification of the C1orf27-binding sites. A , co-IP of α 2A -AR and C1orf27. HEK293 cells were transfected with HA-α 2A -AR together with GFP-C1orf27 and subjected to IP with HA antibodies. B , sequences of the ICL3 and the CT of α 2A -AR. The C1orf27-binding domain in the ICL3 as identified in D is bolded . C , interaction of the ICL3 and the CT of α 2A -AR with C1orf27 in GST fusion protein pulldown assays. D , interactions of different ICL3 fragments with C1orf27 in GST fusion protein pulldown assays. E , summary of progressive deletion to identify the C1orf27-binding domain in the ICL3 of α 2A -AR as shown in D. F , effect of increasing concentrations of NaCl on C1orf27 interaction with S296-L332. G , quantitative data shown in F. H , effect of increasing concentrations of NaCl on C1orf27 interaction with the CT. I , quantitative data shown in H. In each panel, similar results were obtained in at least three separate experiments. Lower panels in C, D, F, and H show GST fusion proteins used in individual experiments. α2A-AR, α2A-adrenergic receptor; Co-IP, co-immunoprecipitation; CT, C terminus; GFP, green fluorescent protein; GST, glutathione S-transferase; HA, hemagglutinin; ICL3, third intracellular loop.

Article Snippet: Antibodies against C1orf27 were purchased from Proteintech.

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Transfection, Immunoprecipitation