Related Experiment Video
Updated: May 31, 2026
14:06
Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays
Published on: November 12, 2012
Secretory pathway genes assessed by high-throughput microscopy and synthetic genetic array analysis
Peter W Bircham1, David R Maass, Christina A Roberts
1Department of Biological Sciences, Victoria University of Wellington, Kelburn Parade, Wellington, New Zealand.
Molecular Biosystems
|July 7, 2011
Summary
The EMC genes, including emc1-6, are crucial for the correct localization of plasma membrane proteins in yeast. Their deletion causes mislocalization to the endoplasmic reticulum, suggesting a role in early secretory pathway transport.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- The precise localization of plasma membrane proteins is essential for cellular function.
- The role of the Essential for Mitotic Growth (EMC) complex in protein transport is not fully understood.
Purpose of the Study:
- To investigate the effect of yeast gene deletions on the localization of the plasma membrane protein Mrh1p-GFP.
- To identify genes involved in the early secretory pathway using a high-throughput screening approach.
Main Methods:
- Developed an automated confocal microscopy procedure for high-content screening.
- Utilized a multi-fluorescence labeling system (Mrh1p-GFP, Redstar2 RFP, mCherry RFP) for cellular compartment visualization.
- Employed image analysis scripts for automated cell segmentation into nucleus, cytoplasm, and cell surface.
Main Results:
- Deletion of EMC genes (emc1Δ-emc6Δ) resulted in the intracellular mislocalization of Mrh1p-GFP to the endoplasmic reticulum.
- CPY processing patterns remained unaffected in these mutants.
- emcΔ hac1Δ double mutants exhibited hypersensitivity to ER-stress, indicating a role for EMC genes in ER homeostasis.
Conclusions:
- The EMC genes play a significant role in the transport of plasma membrane proteins within the early secretory pathway.
- High-content microscopy screening is a valuable tool for discovering functional effects of gene deletions.
- EMC genes are implicated in maintaining endoplasmic reticulum function and responding to stress.
Related Concept Videos
02:46Genetic Screens
Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...
02:34DNA Microarrays
Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...