Environmentally induced foregut remodeling by PHA-4/FoxA and DaF-12/NHR 1Wanyuan Ao, 1Jeb Gaudet, 2W. james Kent, 1Srikanth Muttumu, 1Susan E. Mango 1Huntsman.

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Environmentally induced foregut remodeling by PHA-4/FoxA and DaF-12/NHR 1Wanyuan Ao, 1Jeb Gaudet, 2W. james Kent, 1Srikanth Muttumu, 1Susan E. Mango 1Huntsman cancer institute, University of Utah 2Genome bioinformatics Group, University of California

Pharyngeal devleopment depends on pha-4 Embryos that lack pha-4 fail to generate pharyngeal cells. Many and possibly all genes expressed in the pharynx(at all stages) require pha-4 for activation What causes variation in expression?

Combinatorial regulation Multiple TF’s working towards common or different goals to attain the outcome necessary for tissue differentiation and activity.

1 The Aproach Expression profiles for 339 pharyngeal genes Identified by microarray analysis Five clusters of genes with shared expression profiles were determined for use with topographical expression maps Ph-M (muscle) Ph-MC(marginal cells) Ph-MMC(combine above) Ph-G(glands) Epi (epithelial)

2 Algorithm created to Identify potential cis- regulatory elements Uses set of 194 randomly selected genes not expressed in pharynx as a negative control Seven candidate elements determined

Uncharacterized genes used to validate assignments

Five displayed enhancer activity

Proposed mechanism for expression PHA-4 sites contribute to organ-wide activation throughout pharynx and additional elements impart positional (29-4) cell type (M-2,12-1,12-5) information

3 the M-2 motif Highly selective for pharyngeal muscle. Four genes selected which carried consensus in both C elegans and C briggsae and were expressed in pharyngeal muscle.

M-2 not effected byT-box binding proteins

Yeast one hybrid used to discover transcription factor which binds M-2

Daf-12 is determined a regulatory element in vivo

4 Choice of Growth phase and Dauer development depends on insulin, TGF-B and heterotrimeric guanine nucleotide- binding protein coupled receptor signaling pathways. DAF-12 is affected, what effect does it have on these biological functions?

Down regulation observed in Dauer animals

Food restoration returned animals to growth phase

Daf12 effect repression or loss of activation

PHA-4 binding sites not required for repression