Regulation of signaling genes by TGFbeta during entry into dauer diapause in C. elegans.
BACKGROUND
When resources are scant, C. elegans larvae arrest as long-lived dauers under the control of insulin/IGF- and TGFbeta-related signaling pathways. However, critical questions remain regarding the regulation of this developmental event. How do three dozen insulin-like proteins regulate one tyrosine kinase receptor to control complex events in dauer, metabolism and aging? How are signals from the TGFbeta and insulin/IGF pathways integrated? What gene expression programs do these pathways regulate, and how do they control complex downstream events?


RESULTS
We have identified genes that show different levels of expression in a comparison of wild-type L2 or L3 larvae (non-dauer) to TGFbeta mutants at similar developmental stages undergoing dauer formation. Many insulin/IGF pathway and other known dauer regulatory genes have changes in expression that suggest strong positive feedback by the TGFbeta pathway. In addition, many insulin-like ligand and novel genes with similarity to the extracellular domain of insulin/IGF receptors have altered expression. We have identified a large group of regulated genes with putative binding sites for the FOXO transcription factor, DAF-16. Genes with DAF-16 sites upstream of the transcription start site tend to be upregulated, whereas genes with DAF-16 sites downstream of the coding region tend to be downregulated. Finally, we also see strong regulation of many novel hedgehog- and patched-related genes, hormone biosynthetic genes, cell cycle genes, and other regulatory genes.


CONCLUSIONS
The feedback regulation of insulin/IGF pathway and other dauer genes that we observe would be predicted to amplify signals from the TGFbeta pathway; this amplification may serve to ensure a decisive choice between "dauer" and "non-dauer", even if environmental cues are ambiguous. Up and down regulation of insulin-like ligands and novel genes with similarity to the extracellular domain of insulin/IGF receptors suggests opposing roles for several members of these large gene families. Unlike in adults, most genes with putative DAF-16 binding sites are upregulated during dauer entry, suggesting that DAF-16 has different activity in dauer versus adult metabolism and aging. However, our observation that the position of putative DAF-16 binding sites is correlated with the direction of regulation suggests a novel method of achieving gene-specific regulation from a single pathway. We see evidence of TGFbeta-mediated regulation of several other classes of regulatory genes, and we discuss possible functions of these genes in dauer formation.
Liu
Tao
T
Department of Molecular Biology and Biochemistry, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA. tliu@nel-exchange.rutgers.edu <tliu@nel-exchange.rutgers.edu>
Zimmerman
Karen K
KK
Patterson
Garth I
GI
D000375:Aging / Q000235:genetics / Q000502:physiology; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism / Q000502:physiology; D003577:Cytochrome P-450 Enzyme System / Q000502:physiology; D004268:DNA-Binding Proteins / Q000502:physiology; D029721:Drosophila Proteins / Q000502:physiology; D025461:Feedback, Physiological / Q000502:physiology; D051858:Forkhead Transcription Factors; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D017238:Genes, Helminth / Q000502:physiology*; D018816:Genes, cdc / Q000502:physiology; D053823:Hedgehog Proteins; D007328:Insulin / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D011972:Receptor, Insulin / Q000502:physiology; D011956:Receptors, Cell Surface / Q000502:physiology; D043562:Receptors, G-Protein-Coupled / Q000502:physiology; D012097:Repressor Proteins / Q000502:physiology; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction / Q000502:physiology*; D012903:Smell / Q000235:genetics; D013002:Somatomedins / Q000378:metabolism; D015534:Trans-Activators / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000502:physiology*
D029742:Caenorhabditis elegans Proteins; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D051858:Forkhead Transcription Factors; D053823:Hedgehog Proteins; D007328:Insulin; C452195:LIN-28 protein, C elegans; C095571:LIN-29 protein, C elegans; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D012097:Repressor Proteins; D013002:Somatomedins; D015534:Trans-Activators; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; C108831:daf-16 protein, C elegans; C070844:ptc protein, Drosophila; D003577:Cytochrome P-450 Enzyme System; C107483:DAF-2 protein, C elegans; D011972:Receptor, Insulin