TOR is a structurally and functionally conserved Ser/Thr kinase found in two multiprotein complexes that regulate many cellular processes to control cell growth. Although extensively studied, the localization of TOR is still ambiguous, possibly because endogenous TOR in live cells has not been examined. Here, we examined the localization of green fluorescent protein (GFP) tagged, endogenous TOR1 and TOR2 in live S. cerevisiae cells. A DNA cassette encoding three copies of green fluorescent protein (3XGFP) was inserted in the TOR1 gene (at codon D330) or the TOR2 gene (at codon N321). The TORs were tagged internally because TOR1 or TOR2 tagged at the N or C terminus was not functional. The TOR1D330-3XGFP strain was not hypersensitive to rapamycin, was not cold sensitive, and was not resistant to manganese toxicity caused by the loss of Pmr1, all indications that TOR1-3XGFP was expressed and functional. TOR2-3XGFP was functional, as TOR2 is an essential gene and TOR2N321-3XGFP haploid cells were viable. Thus, TOR1 and TOR2 retain function after the insertion of 748 amino acids in a variable region of their noncatalytic domain. The localization patterns of TOR1-3XGFP and TOR2-3XGFP were documented by imaging of live cells. TOR1-3XGFP was diffusely cytoplasmic and concentrated near the vacuolar membrane. The TOR2-3XGFP signal was cytoplasmic but predominately in dots at the plasma membrane. Thus, TOR1 and TOR2 have distinct localization patterns, consistent with the regulation of cellular processes as part of two different complexes.
- American Society for Microbiology
- TOR1 and TOR2 have distinct locations in live cells
NEC Classifications and Divisions for Hazardous Locations
Electrical equipment in hazardous (classified) locations present a serious threat for workers due to the existence of explosive dust and vapors. Examples of such facilities range from grain mills and gasoline stations to aircraft hangars and chemical manufacturing plants. To prevent devastating explosions that may occur from the ignition of combustible particles in the atmosphere, the National Electric Code (NEC) published guidelines on the proper classification of hazardous locations. Classes, Divisions and...
Hazardous Location LEDs in Chemical Manufacturing Facilities
Employees inside chemical manufacturing plants are exposed to some of the most unforgiving work conditions in the industrial sector. This is because in such environments, workers are exposed to extreme temperatures, dust, volatile chemicals and invisible, explosive gases. Common protection methods against explosive risks include containment, segregation and prevention. Out of all the options, explosive containment is the only method that allows an explosion to take place, in a confined and safe manner through...
Optimal location of artificial recharge of treated wastewater using fuzzy logic approach
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Case study - Remote location requires minimum concept
A leading power supply provider in Austria took on the project of monitoring water levels at a power plant in Montenegro, in two remote locations where no infrastructure is available for installation. On top of that, site visits could only occur every two months, so a device with remote data transmission capabilities was key.Because of these factors, the customer needed an easy solution for their water level monitoring. Long recording time, easy maintenance and the low effort for maintenance were the main reasons...
Case Study - Groundwater: Locating Water Wells
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