Download Challenge Concentration Type Cellular effect NaCl 1.4M Cation

Challenge
NaCl
Concentration Type
1.4M
Cation stress
Galactose
2%
Carbon source
(replaces glucose)
Cellular effect
Extracellular Na+ exposes yeast to
hyperosmosis, and requires intracellular
glycerol accumulation and cell wall and
cytoskeleton strengthening (Hohmann
2002). Na+ enters cells through the K+
transporters Trk1 and Trk2, and
potentially through Pho89 and Nsc1.
Intracellularly, it displaces K+,
challenging cell volume regulation,
intracellular pH and membrane
potentials, protein synthesis, and enzyme
activation (Arino et al. 2010). At acidic
pH, Na+ is exported by Nha1 (Banuelos
et al. 1998). At higher pH, Na+ efflux is
mediated by the Ena proteins, encoded in
a single gene in most natural strains but
in Wine/European strains a different gene
variant introgressed from S. paradoxus
has been amplified into 3-5 similar
paralogs (Warringer et al. 2011). This
introgression/duplication largely defines
natural yeast variation in salt tolerance.
The Ena1 variant plays the critical role in
Na+ tolerance (Haro et al. 1991).
Regulation of salt tolerance genes is
complex, involving the HOG1,
Calcineurin pathway, TOR pathway,
RIM101 and glucose repression
pathways.
Galactose well supports yeast growth as
only energy and carbon source. It enters
cells through the Gal2 permease in a
process that also requires Gal1.
Intracellular galactose is converted into
glucose-1-phosphate by three sequential
reactions that are catalyzed by Gal10,
Gal1 and Gal7 respectively, with Gal10
also being required for re-cycling of a
pathway intermediate. All the galactose
structural
genes
are
coordinately
regulated at the level of transcription in
response to galactose by Gal4p, Gal80p,
and Gal3p (Johnston and Davis 1984;
Lohr et al. 1995). All seven GAL genes
are spatially co-localized in a gene cluster
(Slot and Rokas 2010). Large natural
variations in galactose use is largely
accounted for by loss-of-function
mutations in Gal2, Gal3 (Warringer et al.
Caffeine
2.25mg/mL
Toxin
Rapamycin
0.024µg/ml
Toxin
Phleomycin
2µg/ml
Toxin
Hydroxyurea
2.5mg/ml
Toxin
2011) or loss of the whole GAL cluster
(Slot and Rokas 2010).
A purine, similar to adenine and guanine,
that binds multiple enzymatic targets. In
yeast, caffeine targets the TORC1
complex (Reinke et al. 2006), caffeine
prevents gene conversion by displacing
Rad51 (Tsabar et al. 2015), impairs DNA
repair 26019182, and may interfere with
cell wall generation. Mutations in very
diverse cellular components modulate
caffeine sensitivity. Caffeine trafficking
is not well understood.
A polyketide macrolide produced by
Streptomyces hygroscopicus. Rapamycin
binds the Fpr1 protein (Koltin et al.
1991). This protein-drug complex binds
to and inhibits Tor1 (Heitman et al.
1991), but not Tor2 12408816. Tor1p is a
component of TORC1, a protein complex
regulating nutrient availability and stress
responses (Loewith & Hall 2011).
Mutations in Fpr1 or Tor1 confer
resistance to the drug (Heitman et al.
1991). Loss of non-essential TORC1
components (e.g. Kog1 and Tco089), or
in TORC1 interactors (e.g. Rrd1) confer
rapamycin sensitivity. Rapamycin binds
to the Fpr1 paralog Fpr2 without known
toxic effects 1380159. Rapamycin
trafficking is not well understood.
A 12 component drug complex isolated
from Streptomyces. Binds to DNA,
impedes DNA polymerase and induces
DNA damage and breakdown, potentially
via an oxidative mechanism (Moore,
1988). Arrests cells before entry into Sphase (Nakada et al. 2003) (Weinert et al.
1994). Loss of DNA repair components,
such as Rad6, 9 or 17 cause phleomycin
hypersensitivity (7785323, 8989864).
Phleomycin trafficking is not well
understood.
Hydroxyurea inhibits reduction of
ribonucleotides
to
deoxidized
ribonucleotides (Krakoff et al. 1968), by
binding to and inhibiting the four
component
RNR
(ribonucleotide
reductase) complex. dNTP depletion
impedes synthesis and repair of DNA
Glycine
30mg N/L
Nitrogen source
(replaces
ammonium)
Isoleucine
30mg N/L
Nitrogen source
(replaces
ammonium)
Allantoin
30mg N/L
Nitrogen source
(replaces
ammonium)
(Yao et al. 2003) (Koç et al. 2004),
arresting cells in early S-phase. The RNR
complex consists of four proteins, RNR1,
2, 3, and 4, of which RNR1 and 2 are
essential, and RNR4’s essentiality
depends on the genetic background
(Elledge and Davis, 1987; Elledge and
Davis, 1990; Huang and Elledge, 1997).
Natural variation in hydroxyurea
resistance is partially mediated via the
Hur1 protein (Warringer et al. 2011).
Glycine is generally a very poor nitrogen
source for yeast, but the growth delays,
rates and efficiencies vary between
strains (Ibstedt et al. 2015). Glycine has
no dedicated high affinity permease but is
taken up by the broad specificity amino
acid permeases Gap1 (Jauniaux and
Grenson 1990) and Agp1 and the more
specialized Dip5 and Put4 (Regenberg et
al. 1999). Intracellular glycine is
catabolized into ammonium in three
sequential mitochondrial reactions. These
are catalyzed by a single glycine cleavage
complex with four components: Gcv3,
Gcv1, Gcv2 and Lpd1. Cleavage
reactions requires a folic acid derivative.
Isoleucine is generally a poor nitrogen
source for yeast, but growth delays, rates
and efficiencies vary between strains
(Ibstedt et al. 2015). Isoleucine is take up
by the paralogous high affinity permeases
Bap2 (Grauslund et al. 1995) and Bap3
(Regenberg et al. 1999) as well the low
affinity general amino acid permease
Gap1 (Jauniaux and Grenson 1990).
Isoleucine is catabolized in a single step
reaction to glutamate, using the
mitochondrial Bat1 or the cytoplasmic
Bat2 (Kispal et al. 1996), with Bat1
expressed during exponential growth and
Bat2 in stationary phase.
Allantoin is the primary nitrogen
secretion product of higher mammals,
excluding apes. Yeast utilizes allantoin as
sole nitrogen source, but with varying
delays, rates and efficiencies that largely
maps to variation in Dal4 and Dal1
(Ibstedt et al. 2015). The Dal4 permease
is the only known entrance mechanism
for allantoin. Intracellular allantoin is
catabolized to urea in three sequential
reactions catalyzed by Dal1, Dal2 and
Dal3 respectively (Buckholz and Cooper
1991; Yoo and Cooper 1991a; Yoo and
Cooper 1991b). Urea is converted into
ammonium by the multi-step enzyme
Dur1,2 (Cooper et al. 1980). The
allantoin catabolic genes are regulated by
both general and specific signals that
involve Gln3, Gat1, Dal80, Dal81, and
Dal82 (Magasanik and Kaiser 2002). The
DAL genes are encoded in a tight gene
cluster on Chr IX (Naseeb and Delneri
2012).
Arino J, Ramos J, Sychrova H (2010) Alkali metal cation transport and homeostasis in yeasts
Microbiology and molecular biology reviews : MMBR 74:95-120 doi:10.1128/MMBR.0004209
Banuelos MA, Sychrova H, Bleykasten-Grosshans C, Souciet JL, Potier S (1998) The Nha1 antiporter of
Saccharomyces cerevisiae mediates sodium and potassium efflux Microbiology 144 ( Pt
10):2749-2758
Beese SE, Negishi T, Levin DE (2009) Identification of Positive Regulators of the Yeast Fps1 Glycerol
Channel PLoS genetics 5:e1000738 doi:10.1371/journal.pgen.1000738
Bergstrom A et al. (2014) A High-Definition View of Functional Genetic Variation from Natural Yeast
Genomes Molecular biology and evolution doi:10.1093/molbev/msu037
Bienert GP, Thorsen M, Schussler MD, Nilsson HR, Wagner A, Tamas MJ, Jahn TP (2008) A subgroup
of plant aquaporins facilitate the bi-directional diffusion of As(OH)3 and Sb(OH)3 across
membranes BMC biology 6:26 doi:10.1186/1741-7007-6-26
Buckholz RG, Cooper TG (1991) The allantoinase (DAL1) gene of Saccharomyces cerevisiae Yeast
7:913-923 doi:10.1002/yea.320070903
Cooper TG, Lam C, Turoscy V (1980) Structural analysis of the dur loci in S. cerevisiae: two domains of
a single multifunctional gene Genetics 94:555-580
Grauslund M, Didion T, Kielland-Brandt MC, Andersen HA (1995) BAP2, a gene encoding a permease
for branched-chain amino acids in Saccharomyces cerevisiae Biochimica et biophysica acta
1269:275-280
Haro R, Garciadeblas B, Rodriguez-Navarro A (1991) A novel P-type ATPase from yeast involved in
sodium transport FEBS letters 291:189-191
Hohmann S (2002) Osmotic stress signaling and osmoadaptation in yeasts Microbiology and
molecular biology reviews : MMBR 66:300-372.
Ibstedt S et al. (2015) Concerted evolution of life stage performances signals recent selection on
yeast nitrogen use Molecular biology and evolution 32:153-161 doi:10.1093/molbev/msu285
Jauniaux JC, Grenson M (1990) GAP1, the general amino acid permease gene of Saccharomyces
cerevisiae. Nucleotide sequence, protein similarity with the other bakers yeast amino acid
permeases, and nitrogen catabolite repression European journal of biochemistry / FEBS
190:39-44
Johnston M, Davis RW (1984) Sequences that regulate the divergent GAL1-GAL10 promoter in
Saccharomyces cerevisiae Molecular and cellular biology 4:1440-1448
Kispal G, Steiner H, Court DA, Rolinski B, Lill R (1996) Mitochondrial and cytosolic branched-chain
amino acid transaminases from yeast, homologs of the myc oncogene-regulated Eca39
protein The Journal of biological chemistry 271:24458-24464
Liu Z, Boles E, Rosen BP (2004) Arsenic trioxide uptake by hexose permeases in Saccharomyces
cerevisiae The Journal of biological chemistry 279:17312-17318
doi:10.1074/jbc.M314006200
Lohr D, Venkov P, Zlatanova J (1995) Transcriptional regulation in the yeast GAL gene family: a
complex genetic network Faseb J 9:777-787
Maciaszczyk-Dziubinska E, Migdal I, Migocka M, Bocer T, Wysocki R (2010) The yeast
aquaglyceroporin Fps1p is a bidirectional arsenite channel FEBS letters 584:726-732
doi:10.1016/j.febslet.2009.12.027
Magasanik B, Kaiser CA (2002) Nitrogen regulation in Saccharomyces cerevisiae Gene 290:1-18
Naseeb S, Delneri D (2012) Impact of chromosomal inversions on the yeast DAL cluster PloS one
7:e42022 doi:10.1371/journal.pone.0042022
Regenberg B, During-Olsen L, Kielland-Brandt MC, Holmberg S (1999) Substrate specificity and gene
expression of the amino-acid permeases in Saccharomyces cerevisiae Current genetics
36:317-328
Reinke A, Chen JC, Aronova S, Powers T (2006) Caffeine targets TOR complex I and provides evidence
for a regulatory link between the FRB and kinase domains of Tor1p The Journal of biological
chemistry 281:31616-31626 doi:10.1074/jbc.M603107200
Slot JC, Rokas A (2010) Multiple GAL pathway gene clusters evolved independently and by different
mechanisms in fungi Proceedings of the National Academy of Sciences of the United States of
America 107:10136-10141 doi:10.1073/pnas.0914418107
Talemi SR, Jacobson T, Garla V, Navarrete C, Wagner A, Tamas MJ, Schaber J (2014) Mathematical
modelling of arsenic transport, distribution and detoxification processes in yeast Molecular
microbiology 92:1343-1356 doi:10.1111/mmi.12631
Thorsen M, Jacobson T, Vooijs R, Navarrete C, Bliek T, Schat H, Tamas MJ (2012) Glutathione serves
an extracellular defence function to decrease arsenite accumulation and toxicity in yeast
Molecular microbiology 84:1177-1188 doi:10.1111/j.1365-2958.2012.08085.x
Thorsen M, Lagniel G, Kristiansson E, Junot C, Nerman O, Labarre J, Tamas MJ (2007) Quantitative
transcriptome, proteome, and sulfur metabolite profiling of the Saccharomyces cerevisiae
response to arsenite Physiological genomics 30:35-43
Tsabar M, Mason JM, Chan YL, Bishop DK, Haber JE (2015) Caffeine inhibits gene conversion by
displacing Rad51 from ssDNA Nucleic Acids Res 43:6902-6918 doi:10.1093/nar/gkv525
Warringer J et al. (2011) Trait variation in yeast is defined by population history PLoS genetics
7:e1002111 doi:10.1371/journal.pgen.1002111
PGENETICS-D-10-00228 [pii]
Wieland J, Nitsche AM, Strayle J, Steiner H, Rudolph HK (1995) The PMR2 gene cluster encodes
functionally distinct isoforms of a putative Na+ pump in the yeast plasma membrane The
EMBO journal 14:3870-3882.
Wysocki R, Chery CC, Wawrzycka D, Van Hulle M, Cornelis R, Thevelein JM, Tamas MJ (2001) The
glycerol channel Fps1p mediates the uptake of arsenite and antimonite in Saccharomyces
cerevisiae Molecular microbiology 40:1391-1401
Wysocki R et al. (2004) Transcriptional activation of metalloid tolerance genes in Saccharomyces
cerevisiae requires the AP-1-like proteins Yap1p and Yap8p Molecular biology of the cell
15:2049-2060 doi:10.1091/mbc.E03-04-0236
Wysocki R, Tamas MJ (2010) How Saccharomyces cerevisiae copes with toxic metals and metalloids
FEMS microbiology reviews doi:FMR217 [pii]
10.1111/j.1574-6976.2010.00217.x
Wysocki R, Tamás MJ (2011) Saccharomyces cerevisiae as a model organism for elucidating arsenic
tolerance mechanisms. In: Bánfalvi G (ed) Cellular effects of heavy metal. Springer Verlag,
Heidelberg, pp 87-112
Yoo HS, Cooper TG (1991a) Sequences of two adjacent genes, one (DAL2) encoding allantoicase and
another (DCG1) sensitive to nitrogen-catabolite repression in Saccharomyces cerevisiae Gene
104:55-62
Yoo HS, Cooper TG (1991b) The ureidoglycollate hydrolase (DAL3) gene in Saccharomyces cerevisiae
Yeast 7:693-698 doi:10.1002/yea.320070705