Research
The
main theme of my research at UCI has been the study of transposable
elements (TEs) and the consequences of their activity in the genomes
of plants. TEs are famous for their sometimes dramatic deleterious
effects, but can they also contribute in a more positive manner
to their host’s genome? The answer is an emphatic “yes”:
TEs contribute to protein coding sequence and are able to alter
the expression of genes, making them key players in the evolution
of genomes. Since the sole function of a TE is to copy itself, TEs
are surprisingly rare in genomes. What are the forces keeping TEs
from overwhelming genomes? I am interested in how the forces of
recombination and selection keep TEs under control.
I
have discovered numerous genes in Arabidopsis thaliana
where TE sequence is present in the protein coding sequence. Using
stringent BLAST searches, I have determined that a large number
of these genes containing TE sequence are also expressed. This bioinformatic
study used experimentally-derived Expressed Sequence Tag (EST) sequences,
to confirm the TE-gene chimaera’s expression and to add a
robust layer of “real-world” data. However, since TEs
can sometimes pick up gene fragments, as well as insert into genes,
I used the phylogenetic histories of gene families in the Arabidopsis
genome to determine the direction of sequence transmission and to
infer with confidence whether an actual insertion event took place
or not. This paper is currently in preparation and will be submitted
this summer.
The
model plant Arabidopsis thaliana and its close relative
Arabidopsis lyrata possess different mating systems. The
former is highly selfing, while the latter is an obligate outcrosser.
Population genetics theory predicts that recombination rates are
lower in selfers compared to outcrossers and that selection is weaker
in areas of low recombination. TE copy number can be maintained
in genomes by either purifying selection acting against deleterious
insertions, or by selection acting against harmful non-homologous
recombination. Using a method called Transposon-Insertion Display
(TID) I have gathered a large TE insertion frequency dataset for
eight different TE families across both species, in 47 ecotypes
per species. Using these data I will be able to tease apart the
relative contributions of selection and recombination in controlling
TEs in plant genomes, hence shedding more light on the structure
and evolution of plant genomes themselves.
My
final project of my Ph.D. thesis studies the effect of TE insertion
on gene expression. TE polymorphism data was gathered for 22 genes
from 48 A. thaliana ecotypes where the TE had inserted
close to a gene (< 300 b.p.). One would hypothesise if a TE (often
with its own promoters) landed in the regulatory region of a gene
it would have an effect upon the expression of that gene. I intend
to investigate this by producing F1 hybrids that contain one allele
with the TE absent, and another with the nearby TE present. I will
then measure the relative expression of both alleles. Using F1 hybrids
will allow me to infer cis-regulatory change, as both alleles
will exist in the same trans-factor background, which would be consistent
with the hypothesis that TE insertion in promoters results in cis-regulatory
change.
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