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Profiles
Eva
Nokes
Graduate
Student
enokes
[at] brandeis.edu
The ability
of C. elegans to sense multiple chemical signals in their
environment is mediated by individual chemosensory neuron subtypes,
each of which senses a unique subset of chemicals, and expresses
a defined subset of chemoreceptor and other signaling genes. We
are interested in understanding the transcriptional rules that govern
the expression of chemosensory genes specifically in individual
sensory neuron subtypes. We selected the AWB neurons for our study
since we have identified a set of AWB-selective chemoreceptors and
other genes via expression profiling from isolated AWB neurons (Colosimo
et al., 2004).
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| a)
The promoter of sru-38 fused to GFP shows expression in the
AWB as well as ASH pair of chemosensory neurons. b) When four
base pairs in the same GFP fusion are mutated, GFP expression
in lost specifically in the AWB neurons. |
Using deletions
and point mutations, I have now identified cis-regulatory motifs
driving AWB-specific expression in the promoters of several chemoreceptor
genes. Interestingly, these motifs share only a 5 base pair core
sequence, while the required flanking sequences are distinct for
each gene. To determine whether genes other than chemoreceptor genes
use similar regulatory mechanisms, I have also defined minimal equences
required for the expression of more broadly expressed genes such
as odr-4 and kap-1. The LIM homeodomain transcription
factor LIM-4 has previously been shown to regulate its own expression
in the AWB neurons (Sagasti
et al, 1999), and to regulate the expression of all known
AWB-specific genes. I am also identifying the sequences in the lim-4
promoter required for autoregulation, and determining whether LIM-4
directly regulates expression of the identified chemoreceptor genes
using in vitro and in vivo binding assays. My preliminary data suggest
that the cis-regulatory sequences of the studied chemoreceptor gene
orthologs in C. briggsae are highly divergent, suggesting
independent divergence of chemoreceptor gene regulatory and protein
coding sequences. I am also performing forward genetic screens to
identify additional trans-acting factors that drive gene expression
in the AWB neurons.
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