INTERACTIONS BETWEEN DREB1A/CBF3

AND HAC1 IN RESPONSE TO COLD STRESS

Julia Proctor
Background/Hypothesis
Due to global climate change, weather is
increasingly unpredictable, which impacts plant
productivity3. Due to their sessile nature, plants are
unable to escape environmental stresses and must
be able to acclimate3.
HAC1 is a histone acetyltransferase protein in
Arabidopsis thaliana, responsible for mediating
DNA transformations to upregulate or suppress
transcription of stress-response genes.
Our hypothesis is that DREB1a, a transcription
factor, recruits the histone acetyltransferase HAC1
to upregulate the transcription of COR78, a coldresponse gene, under cold stress in Arabidopsis
thaliana1. Having the ability to manipulate this
protein family may serve as an alternative approach
to increasing crop yields in the future2.
Methods
Grow 5 WT (CS60000) and 5 MT
(SALK_080380) Arabidopsis thaliana plants for 21
days. Leave 1 WT and 1 MT in the standard
greenhouse conditions for control against our
experimental conditions. The rest of the plants will
be placed in boxes with timed lights in either cold
(4C) or room (26C) temperature treatment.
Each of the remaining pairs (1 WT, 1 MT) will
start in 26C and be transferred to 4C at varying
treatment time points (0, 2, 6, and 24 hours,
respectively).
Isolate RNA from ~100mg of leaf tissue from
each plant. Perform RT-PCR to compare expression
of HAC1, DREB1A, COR78, and UBQ5. Run a gel
to assess the relative quantities of PCR product for
each sample.
Observations/Results

Figure 1: Diagram of our proposed mechanism in how

DREB1a is involved with cold-stress response.

Figure 2: Gels showing gene expression of HAC1, UBQ5,

COR78 and DREB1a. Labeled lanes 1-5 represent WT
plants, lanes 6-10 represent MT, and lanes 11 represent a no
template (-T). Lane 12 is only present for UBQ5 and was a
-RT control. Treatments: no treatment (NT); 0h, 2h, 6h, and
24h represent hours exposed to cold stress (4C)

Conclusion/Discussion
Without the UBQ5 gel showing relatively equal
expression levels, there are limitations in our ability
to assess our data. The best explanation for the (lack
of) UBQ5 data is due to primer error.
However, if we evaluate our data as is, we see
that HAC1 expression is present in all WT samples,
but not in MT samples (except for the no treatment
lane). This reinforces that the mutant plants that we
used had a T-DNA insert that disrupted the HAC1
gene. It appears that HAC1 is not recruited in
response to cold stress since the levels drop as
exposure to the cold stress increases.
A possible explanation for why the no
treatment MT shows HAC1 expression is that it was
not truly a MT plant. All of our plants except for the
no treatment plants were genotyped. Since the data
on the MT plant with no treatment is contradictory,
the reliability of any of the data collected using this
plant may be questionable.
COR78 showed no difference in expression
levels regardless of plant type or treatment. This
may be because the PCR was run to saturation.
DREB1a expression starts low and shows an
increase as the exposure to the cold treatment
increases. This may be due to the effects of
acclimation. The similar expression levels between
WT and MT plants indicate HAC1 is not recruited,
but that DREB1a does seem to respond to the cold
stress stimulus.
References
Ahmadizadeh, (2014) Biharean Biologist. 8:2.

Akhtar, (2012) Journal of Genetics. 91:3.

Oakenfull, (2013) PLOS ONE. 8:1.