Mapping of an internal protease cleavage site in the Ssy5p component of the amino acid sensor of Saccharomyces cerevisiae and functional characterization of the resulting pro- and protease domains by gain-of-function genetics

Eukaryotic Cell
Peter PoulsenMorten C Kielland-Brandt

Abstract

Ssy5p is a 77-kDa protein believed to be a component of the SPS amino acid sensor complex in the plasma membrane of Saccharomyces cerevisiae. Ssy5p has been suggested to be a chymotrypsin-like serine protease that activates the transcription factor Stp1p upon exposure of the yeast to extracellular amino acid. Here we overexpressed and partially purified Ssy5p to improve our understanding of its structure and function. Antibodies against Ssy5p expressed in Escherichia coli were isolated and used to detect Ssy5p processing in S. cerevisiae cells. Partial purification and N-terminal sequencing of processed Ssy5p revealed in vivo cleavage of Ssy5p between amino acids 381 and 382. We also isolated constitutively signaling SSY5 mutants and quantified target promoter activation and Stp1p processing. One mutant contained an amino acid substitution in the prodomain, whereas three others harbored amino acid substitutions in the protease domain. Dose-response analysis indicated that all four mutants exhibited increased basal levels of Stp1p processing. Interestingly, whereas the three constitutive mutants mapping to the protease domain of Ssy5p exhibited the decreased 50% effective concentration (EC(50)) characteristic of constitutive mut...Continue Reading

References

Jan 1, 1991·Methods in Enzymology·F Sherman
Nov 30, 1995·Biochimica Et Biophysica Acta·M GrauslundH A Andersen
Dec 1, 1993·Current Opinion in Cell Biology·D BakerD A Agard
Nov 10, 1998·Nature Structural Biology·N K SauterD A Agard
Oct 3, 1999·Biotechnology and Bioengineering·G D DavisR G Harrison
Jun 22, 2000·Journal of Molecular Biology·L A KelleyM J Sternberg
Feb 24, 2001·Molecular & General Genetics : MGG·P S NielsenH A Andersen
Dec 28, 2002·Genes & Development·Claes Andréasson, Per O Ljungdahl
Sep 29, 2004·Trends in Biochemical Sciences·Inge HolsbeeksJohan M Thevelein
Oct 1, 2004·Molecular and Cellular Biology·Nathalie SpielewoyCurt Wittenberg
Jan 26, 2005·Biochemical Society Transactions·P PoulsenM C Kielland-Brandt

❮ Previous
Next ❯

Citations

Nov 27, 2008·Proceedings of the National Academy of Sciences of the United States of America·Cheol-Sang Hwang, Alexander Varshavsky
Dec 4, 2010·The Journal of Biological Chemistry·Sylvester TumusiimeZhengchang Liu
Jun 10, 2011·Molecular Biology of the Cell·Deike J OmnusPer O Ljungdahl
Jun 17, 2006·Genes & Development·Claes AndréassonPer O Ljungdahl
Apr 28, 2010·Molecular and Cellular Biology·Thorsten PfirrmannPer O Ljungdahl
Jan 23, 2009·American Journal of Physiology. Endocrinology and Metabolism·Harinder S Hundal, Peter M Taylor
Jun 12, 2013·PloS One·Thorsten PfirrmannThomas Hollemann
Mar 16, 2012·Genetics·Per O Ljungdahl, Bertrand Daignan-Fornier
Mar 1, 2008·Molecular Membrane Biology·Peter PoulsenMorten C Kielland-Brandt
Sep 13, 2015·Critical Reviews in Biochemistry and Molecular Biology·Kunio NakatsukasaTakumi Kamura
Jun 17, 2016·Molecular Biology of the Cell·Laura MelladoEduardo A Espeso
Feb 29, 2008·Annual Review of Genetics·Shadia ZamanJames R Broach
Apr 18, 2018·The Journal of Biological Chemistry·António MartinsPer O Ljungdahl
Jan 18, 2017·The EMBO Journal·Asier González, Michael N Hall
Aug 24, 2017·Scientific Reports·Anna MuszewskaKrzysztof Ginalski

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.