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
Case Study - Groundwater: Locating Water Wells
South Sudan: 4000 L/min water wellDate: February 25, 2015Site: Yeri Mozo Village, South Sudan, AfricaEquipment: SuperStingR8 WiFi, SwitchBox56 and passive cables spaced at 5m.Software: EarthImager2D with a finite element inversion model of dipole-dipole array data.Results: Resistivity imaging successfully located a very high production potable water well with 4000 liter/hour static yield.
Simultaneous nitrification, denitrification and phosphorus removal (SNDPR) in a full-scale water ...
The combination of simultaneous nitrification-denitrification (SND) with enhanced biological phosphorus removal (EBPR) provides a more efficient and economically viable option for nutrient removal from municipal wastewater compared to conventional two-step nitrification-denitrification. This study analyzed the nutrients (N and P) profiles in a full-scale municipal wastewater reclamation plant (WRP) located in the tropical region, in which more than 90% of nitrogen was removed. Interestingly, average SND efficiency...
Antibiotics in groundwater under locations with high livestock density in Germany
Antibiotics are deployed in large quantities both in human and in veterinary medicine. Studies show that antibiotic residues occur in the environment (e.g. soil and surface waters). In some cases they were also detected in ground and drinking water. However, the degree of groundwater pollution by antibiotic residues from livestock farming is unknown. Therefore, the federal environment agency (UBA) supported a project that aimed to investigate near-surface groundwater samples in regions of high livestock density...
Locating Leaks for Cooling Water Systems
This article illustrates how to locate leaks on cooling water systems in loud, industrial environments like Nuclear or Coal power generating plants without digging or putting anything inside the pipeline. Infrared-Energy Pattern Analysis (IR-EPA) works on all pipeline sizes and all pipeline materials because it identifies the water outside the pipeline instead of listening for leak noises like traditional leak detection techniques. IR-EPA is safe due to its passive nature and only monitors the ground surface for...
Typical Sensor Location Considerations
The sensor mounting location within a space may be dependent on many variables. There are no specific formulas since each application will have it’s own determining factors and priorities. These include, but are not limited to: source of gas: Is the source a combustion exhaust from a vehicle or furnace or a vapour from a tank spill, pipe leak, or faulty valve? density of gas: some gases are light than air and will rise, others are heavier and will migrate toward to the floor or into a pit use of the...