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
BRIMS.ca Goes Live
Alberta’s Bio-Resource Information Management System (BRIMS) was formally launched on January 24, 2018, in Edmonton, Alberta. Key representatives from the Government of Alberta, the University of Alberta, Alberta Innovates and other important industries gathered in the Silvacom head office for the public unveiling and live demonstration of the BRIMS web application. With an ever-expanding demand for green or sustainable products in Alberta, BRIMS is geared to help propel investments in the bio-economy to...
How are Green Buildings Built? A Closer Look at a Modern Marvel
Green building refers to an environment friendly structure which is built in such a manner that it continues to be environmentally responsible and resource efficient throughout its life. When we say throughout its life, it includes everything right from an eco-friendly design to the use of material which will make sure that the environment is not harmed even when the structure in question is demolished. The concept of green architecture has come a long way, with modern green buildings being built by taking into...
EEA Signals 2015 - Living in a changing climate
Our climate is changing. Scientific evidence shows that the global average temperature is rising, and rainfall patterns are shifting. It also shows that glaciers, Arctic sea ice and the Greenland ice sheet are melting. The Intergovernmental Panel on Climate Change′s Fifth Assessment Report shows that the warming since the mid-20th century is predominantly due to an increase in greenhouse-gas concentrations as a result of emissions from human activities. Combustion of fossil fuels and changes in land use are...
Living with the floods, not against them
“Yes, we are illiterate sir, but we are engineers as far as our experience with water and rivers is concerned”, says Phushi Mahato, a villager in the Gosi Kandak floodplains in North-Bihar in India. As an outcome of water resource management that failed to understand the role of fluvial regimes in delta, Phushi and his fellow village members have now become vulnerable to floods vulnerable while in the past they used to live with the natural flows, with their farming and fisheries fuelled by dynamic...
Emerging Technologies – How will they change our lives?
Forward planning by governmental agencies is often criticized for being too focused on short term political mandate priorities. A recent report by Policy Horizons Canada, a federal government agency, on emerging technologies is a refreshing change from that norm. The report explored how emerging technologies will shape the economy and our society, and what challenges (and opportunities) they will create. The Horizons report does not predict the future. Rather, it systematically explores a range of futures...