NDLI: Secretory pathway genes assessed by high-throughput microscopy and synthetic genetic array analysis

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Contents list

Regulation of integrins by conformation and traffic: it takes two to tango

Global risk transformative prioritization for prostate cancer candidate genes in molecular networks

Photochemical control of bacterial signal processing using a light-activated erythromycin

A novel β-loop scaffold of phage-displayed peptides for highly specific affinities

Characterization of AziR, a resistance protein of the DNA cross-linking agent azinomycin B

MicroRNAs dysregulated in breast cancer preferentially target key oncogenic pathways

A metabolic system-wide characterisation of the pig: a model for human physiology

Secretory pathway genes assessed by high-throughput microscopy and synthetic genetic array analysis

Small delay, big waves: a minimal delayed negative feedback model captures Escherichia coli single cell SOS kinetics

Serum metabolic profiling and features of papillary thyroid carcinoma and nodular goiter

DIPOS: database of interacting proteins in Oryza sativa

Modeling synergistic drug inhibition of Mycobacterium tuberculosis growth in murine macrophages

Proteomic responses in Arabidopsis thaliana seedlings treated with ethylene

Complexome of Escherichia coli cytosolic proteins under normal native conditions

From Cerius² based stereoview to mouse and enzyme: the model systems for discovery of novel urokinase inhibitors

PELDOR analysis of enzyme-induced structural changes in damaged DNA duplexes

pH-responsive, DNA-directed reversible assembly of graphene oxide

Identification of cytoplasmic and membrane-associated complexes in human embryonic stem cells using blue native PAGE

In silico pharmacology suggests ginger extracts may reduce stroke risks

Traditional Chinese medicine, a solution for reducing dual stroke risk factors at once?

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Secretory pathway genes assessed by high-throughput microscopy and synthetic genetic array analysis

Content Provider	Royal Society of Chemistry (RSC)
Author	Bircham, Peter W. Atkinson, Paul H. Maass, David R. Kiew, Poh Y. Roberts, Christina A. Yegambaram, Manivannan Jack, Cameron A. Low, Yee S. Matthews, James
Copyright Year	2011
Abstract	We developed a procedure for automated confocal microscopy to image the effect of the non-essential yeast gene deletion set on the localisation of the plasma membrane GFP-labelled protein Mrh1p-GFP. To achieve this it was necessary to devise an expression system expressing Redstar2 RFP-fluorescence specifically in the nucleus, mCherry RFP at a lower intensity in the cytoplasm and Mrh1p-GFP in the plasma membrane. This fluorescence labelling scheme utilising specifically designed image analysis scripts allowed automated segmentation of the cells into sub-regions comprising nuclei, cytoplasm and cell-surface. From this high-throughput high content screening approach we were able to determine that gene deletions including emc1Δ, emc2Δ, emc3Δ, emc4Δ, emc5Δ and emc6Δ, caused intracellular mislocalisation at the ER of a plasma membrane protein Mrh1p-GFP. CPY processing patterns were unaffected in these mutants and collectively our data suggest a transport role for the EMC genes within the early secretory pathway. HAC1 is central to the unfolded protein response (UPR) and in its absence, i.e. the absence of UPR, emc1Δ-, emc3Δ-, emc4Δ-, emc5Δ–hac1Δ double mutants were specifically hypersensitive to ER-stress (tunicamycin) lending credence to the usefulness of the high content microscope screening for discovery of functional effects of single mutants.
Starting Page	2589
Ending Page	2598
Page Count	10
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	7
Issue Number	9
Journal	Molecular BioSystems
DOI	10.1039/c1mb05175j
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Membrane protein Cell membrane Secretion Tunicamycin Confocal microscopy Fluorescence microscope Gene expression Protein Dell EMC Unfolded protein response Gene Microscopy UPR EMC MCherry Saccharomyces cerevisiae Endoplasmic reticulum Cytoplasm
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Endoplasmic reticulum-associated degradation (ERAD) and unfolded protein response (UPR): genetic studies in Saccharomyces cerevisiae

Cell-free sorting of peroxisomal membrane proteins from the endoplasmic reticulum.

The unfolded protein response regulates multiple aspects of secretory and membrane protein biogenesis and endoplasmic reticulum quality control (2000)

Effects of inactivation and constitutive expression of the unfolded- protein response pathway on protein production in the yeast Saccharomyces cerevisiae.

The Unfolded Protein Response Is Required for Haploid Tolerance in Yeast

Signal sequences specify the targeting route to the endoplasmic reticulum membrane (1996)

Activation of mammalian unfolded protein response is compatible with the quality control system operating in the endoplasmic reticulum.

The Unfolded Protein Response: No Longer Just a Special Teams Player

Mutant in three novel complementation groups inhibit membrane protein insertion into and soluble protein translocation across the endoplasmic reticulum membrane of Saccharomyces cerevisiae (1992)

Secretory pathway genes assessed by high-throughput microscopy and synthetic genetic array analysis

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Endoplasmic reticulum-associated degradation (ERAD) and unfolded protein response (UPR): genetic studies in Saccharomyces cerevisiae

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Effects of inactivation and constitutive expression of the unfolded- protein response pathway on protein production in the yeast Saccharomyces cerevisiae.

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Secretory pathway genes assessed by high-throughput microscopy and synthetic genetic array analysis