Chick Hairy1 protein interacts with Sap18, a component of the Sin3/HDAC transcriptional repressor complex.
BACKGROUND
The vertebrate adult axial skeleton, trunk and limb skeletal muscles and dermis of the back all arise from early embryonic structures called somites. Somites are symmetrically positioned flanking the embryo axial structures (neural tube and notochord) and are periodically formed in a anterior-posterior direction from the presomitic mesoderm. The time required to form a somite pair is constant and species-specific. This extraordinary periodicity is proposed to depend on an underlying somitogenesis molecular clock, firstly evidenced by the cyclic expression of the chick hairy1 gene in the unsegmented presomitic mesoderm with a 90 min periodicity, corresponding to the time required to form a somite pair in the chick embryo. The number of hairy1 oscillations at any given moment is proposed to provide the cell with both temporal and positional information along the embryo's anterior-posterior axis. Nevertheless, how this is accomplished and what biological processes are involved is still unknown. Aiming at understanding the molecular events triggered by the somitogenesis clock Hairy1 protein, we have employed the yeast two-hybrid system to identify Hairy1 interaction partners.


RESULTS
Sap18, an adaptor molecule of the Sin3/HDAC transcriptional repressor complex, was found to interact with the C-terminal portion of the Hairy1 protein in a yeast two-hybrid assay and the Hairy1/Sap18 interaction was independently confirmed by co-immunoprecipitation experiments. We have characterized the expression patterns of both sap18 and sin3a genes during chick embryo development, using in situ hybridization experiments. We found that both sap18 and sin3a expression patterns co-localize in vivo with hairy1 expression domains in chick rostral presomitic mesoderm and caudal region of somites.


CONCLUSIONS
Hairy1 belongs to the hairy-enhancer-of-split family of transcriptional repressor proteins. Our results indicate that during chick somitogenesis Hairy1 may mediate gene transcriptional repression by recruiting the Sin3/HDAC complex, through a direct interaction with the Sap18 adaptor molecule. Moreover, since sap18 and sin3a are not expressed in the PSM territory where hairy1 presents cyclic expression, our study strongly points to different roles for Hairy1 throughout the PSM and in the prospective somite and caudal region of already formed somites.
Sheeba
Caroline J
CJ
Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal. sheeba@ecsaude.uminho.pt <sheeba@ecsaude.uminho.pt>
Palmeirim
Isabel
I
Andrade
Raquel P
RP
D000818:Animals; D030161:Avian Proteins / Q000235:genetics / Q000378:metabolism*; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D001683:Biological Clocks / Q000502:physiology; D019521:Body Patterning; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D056264:Sin3 Histone Deacetylase and Corepressor Complex; D019170:Somites / Q000502:physiology*; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
D030161:Avian Proteins; D051792:Basic Helix-Loop-Helix Proteins; D002352:Carrier Proteins; D006655:Histone Deacetylases; D056264:Sin3 Histone Deacetylase and Corepressor Complex; C109424:hairy1 protein, Gallus gallus