Gene expression of Saccharomyces cerevisiae cells exposed to commercial wood preservatives by Microa

1
Gene expression of Saccharomyces cerevisiae cells
exposed to commercial wood preservatives by
Microarray Analysis

• Madison M. Stevens,1 Gary P. Lutz,2 Consuelo J.
  Alvarez.1 1Department of Biological and
  Environmental Sciences, 2Prince Edward County
  High School, Longwood University, Farmville,
  Virginia

RESULTS.
Tables 1 (top) and 2 (bottom) Genes highlighted
are a subset of induced (red) and repressed
(green) genes involved in the cell division
cycle. A subset of unknown genes are highlighted
in blue
Figs. 6 (on left) A subset of induced and
repressed genes with a foreground of red and
green dye intensities equal to or above 200 and a
gene expression fold change of 2 or greater
YGL239C Score6881 bits (3471),Expect0.0
Identities3600/3602 (99), Gaps 0/3602 (0)
StrandPlus/Plus Query 163 AATCCTGTGGACTTT
..GAGTAAGC 3764



Sbjct 1 AATCCTGTGGACTTT
..GAGTAAGC 3602
YKL071W Score8407 bits (4241),Expect0.0
Identities4328/4328 (100), Gaps0/4328 (0)
StrandPlus/Plus Query 1 AAGCTTCCTCATTTCGT
..AGAATCTATTTCATA 4328


Sbjct 6785 AAGCTTCCTCATTTCGT.
. AGAATCTATTTCATA 11112
Fig. 7 A schematic drawing showing where some
induced (red) and repressed (green) genes are
located in the budding yeast cell, Saccharomyces
cerevisisae
Fig. 8 Blast results for 2 genes of unknown
function
CONCLUSIONS. The data that has been collected to
date in these experiments provide interesting
areas for evaluation. Previous investigators
using these wood preservatives have indicated
that these chemicals are carcinogenic, so we
expect to see induction and repression of genes
involved in the cell cycle division. With the
creosote data, there are indications that the
genes that we have identified as being repressed
are implicated in mismatch DNA repair, signal
transduction during conjugation of cellular
fusion, bud neck formation, and DNA-directed RNA
polymerase (see YDR097C, YDR461W, YDR507C,
YFL036W genes highlighted in green in table 1and
in figure 7). In similar manner, there are
indications that the genes that we have
identified as being induced are involved in cell
wall organization and biogenesis, bud tip
formation, bud neck formation, and stress
response (see YLO105C, YDR309C, YOL007C,
YOL052C-A genes highlighted in red in table 1and
in figure 7). With the pentachlorophenol data,
there are indications that the genes that we have
identified as being repressed are involved in
ribosomal large subunit biogenesis and assembly,
purine base metabolism, pentose-phosphate shunt,
chromatin modeling, and telomere maintenance (see
YJL122W, YKR081C, YGL234W, YOR095C, YFR037C,
YLR150W genes highlighted in green in table 2
and in figure 7). In a similar manner, there are
indications that the genes that we have
identified as being induced are concerned with
mitochondrial organization and biogenesis, signal
transduction, and response to drugs (see YJL116C,
YOR371C, YDR011W genes highlighted in red in
table 2 and in figure 7). Additionally,
blastn analysis of some genes of unknown
function (biological, molecular, and cellular)
indicate that there are similarities between the
unknown genes and genes involved in cell
division cycle (YGL239C, YLR154W-A) or genes
involved in stress response like YKL071W (see
figure 8). Lastly, the pentachlorophenol data
indicate that one third of the genes effected
during the exposure to this chemical are involved
in protein biosynthesis (14 out of 44).
Fig. 3B Close up of PtCP microarray chip
Fig. 5 Creosote expression file in Microsoft excel
Fig. 3A Scanned PtCP microarray chip
Fig. 4 Gene expression color scale
FUTURE WORK. In order to validate the results
obtained in these microarray experiments, real
time-polymerase chain reaction (RT-PCR)
experiments performed with the highly induced or
highly repressed genes are planned. For genes
with currently unknown cellular function,
cellular localization will be determined by
fluorescence after gene fusion of green
fluorescent protein to the respective gene. In
addition, sequences of unknown genes will be
evaluated using the blastn and blastp
programs to check for potential domains that
could provide indications of their biological and
molecular function.

• REFERENCES.
• Agency for Toxic Substances and Disease
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ACKNOWLEDGMENTS. Longwood Foundation and CAS
Deans Funds, Longwood Cormier Citizen
Scholarship, Genomic Consortium for Active
Teaching (GCAT) and its members, and a special
acknowledgment to Dr. Leigh Lunsford, Dr. Phillip
Poplin and Ms. Ashley Swandby in the Department
of Mathematics and Computer Science.