Stress Responses & Gene Expression

• plants must
adapt to
stresses
because of
their
sedentary
lifestyle

Fig. 22.2, Buchanan et al.

 Adaptation and Acclimation
• Adaptation - evolutionary changes that
enable an organism to exploit a certain
niche. These include modification of existing
genes, as well as gain/loss of genes.
– e.g., thermophilic enzymes in organisms that tolerate
high temperature
• Acclimation – inducible response(s) that allows
an organism to tolerate an unfavorable or lethal
(to some plants) change in the environment.
– e.g., heat shock response
 Types of Stress
Abiotic
1. heat
2. cold
3. drought
4. salt
5. wind
6. oxidative
7. anaerobic
8. heavy metals
9. wounding
10. nutrient deprivation
11. excessive light
Biotic
12. pathogens
13. herbivores

Focus on heat, cold, anaerobiosis, oxidative, and biotic stress

Plants respond to stresses as individual cells and as whole organisms. Stress-
induced signals can be transmitted throughout plant, making other parts more
ready to withstand stress.
Heat Stress (or Heat Shock) Response

• Discovered in Drosophila, polytene chromosome

puffing
• Specific response to temperatures ~10-15oC
above normal growth temp.
• Ubiquitous
• Conserved
• Rapid
• Transient
• Dramatic change in pattern of protein synthesis
Heat stress effects on protein synthesis in soybean seedlings (J. Key).
Heat
stress/shock
protein synthesis
in the
cyanobacterium
Synechococcus.
Generalized order of events in heat
shock response
Initial events:
1. Inhibition of protein synthesis
2. Inhibition of transcription & RNA
processing
3. Induction of new hs (hsp) mRNAs
4. Pre-existing cellular mRNAs still present
but not translated (initiation factor
eIF4A & B deactivated)
 Phase II events
5. Partial restoration of protein synthesis,
mainly translation of hsp mRNAs
6. Accumulation of hsp (heat shock
proteins)
7. Gradual resumption of normal cellular
protein synthesis
8. Decline in hsp synthesis
 Function: Thermotolerance
• Enables organisms to survive high temp.
• A sub-lethal heat shock allows organisms to
survive a lethal treatment.
• Production of hsps correlate with acquired
thermotolerance.
• Some mutants (yeast) and transgenic plants with
altered expression of certain hsps don’t show
thermotolerance.
 28oC 40oC  45oC 45oC

Soybean seedlings.
Fig. 22.42, Buchanan et al.
 Heat Shock Proteins (hsp)
• ~100, ~90, ~70, and ~60 kDa

• Low molecular weight (LMW) hsp: ~27, ~20-22,

~15-18 kDa

• all induced within 30 min.

• more LMW hsp in plants

• 2-Dimensional gel electrophoresis and

molecular cloning indicates most hsps are
families of related proteins, particularly hsp70
and the LMW hsps
 HSP functions
• 1 LMW hsp is ubiquitin, which tags
proteins to be degraded by the
proteasome
• hsp90, hsp70, and hsp60 involved in
protein folding: "molecular chaperones"
• hsp100 may promote translation of hsp
mRNAs, via CAP-independent
mechanism
 HSP70, a chaperonin
• Essential gene
• Homologues found in cytoplasm, ER lumen,
mitochondria, and chloroplasts
• function in protein targeting and assembly in normal
(non-stressed) cells, hydrolyze ATP
• Constitutive & heat-induced (cytoplasmic) forms
– the heat-induced form first appears in the nucleolus,
then goes to cytoplasm (may protect pre-ribosomes
from heat stress?)
• Also, some hsp70s are light-induced; chloroplast hsp70
helps protect PSII from light/heat damage in Chlamy
 HSP60 (cpn60)
• first one termed "molecular chaperone“
• Discovered as the RuBPCase LS binding protein
that participates in RuBPCase assembly
• in eucaryotes, only in mitochondria and plastids
• similar to E. coli GroEL gene
• exists as abundant 720 kD complex with two
subunits of 61 and 60 kDa (ATPase)
• associates with cpn10 (GroES homologue)
• facilitates folding/ assembly of other proteins
 LMW HSPs
• highly heat-induced
• 4 nuclear gene families:
1. Class I cytoplasmic
2. Class II cytoplasmic
3. Chloroplast localized
4. Endomembrane localized (ER)
• found in organelles only in plants
• function mostly unknown
• aggregate in vivo into "heat shock granules"
 HSP regulation
• most work on LMW hsp in plants
• induction is mainly transcriptional but
also translational control (hsp mRNAs
preferentially translated)
• genes induced coordinately, but not
equally in all tissues
• light can also induce some LMW hsps
Soybean Hsp
mRNAs that are
strongly
induced.

Temp. oC
 Cis-acting transcriptional regulatory
elements
• HSE (heat shock elements) in the 5' regions:
– ~10-15 bp partial palindromes
– multiple copies required
– also found in other HS genes (e.g., hsp 70)
– similar to HSEs in animals
HSEs in
plants
and
animals.
 Heat-shock transcription factor (HSF)

• studied mostly in animals and yeast

• Binds to HSEs
• Contains leucine zipper motifs
• Binds DNA as a trimer
• Activity is induced by heat, and
phosphorylation
• Activity Repressed by HSP70
Fig. 22.43, Buchanan et al.
Fig. 22.44, Buchanan et al.