Developmentally regulated synthesis of p8, a stress-associated transcription cofactor, in diapause-destined embryos of Artemia franciscana

Diapause-destined embryos of the crustacean Artemia franciscana arrest as gastrulae, acquire extreme stress tolerance, and enter profound metabolic dormancy. Among genes upregulated at 2 days postfertilization in these embryos is a homologue of p8, a stress-inducible transcription cofactor. Artemia p8 is smaller than vertebrate homologues but shares a basic helix-loop-helix domain and a bipartite nuclear localization signal. Probing of restriction digested DNA on Southern blots indicated a single Artemia p8 gene and 5′-RACE specified 2 transcription start sites. Several putative cis-acting regulatory sequences, including two heat shock elements, appeared upstream of the p8 transcription start site. Artemia p8 mRNA increased sharply at 1 day postfertilization in diapause-destined embryos and then declined, whereas p8 protein appeared 2 days postfertilization and remained relatively constant throughout development, indicating a stable protein. p8 was not detectable in nauplius-destined (nondiapause) Artemia embryos. Immunofluorescent staining revealed p8 within Artemia nuclei. The results support the idea that p8, a known stressresponsive transcription cofactor, mediates gene expression in diapause-destined Artemia embryos. p8 is the first diapause-related transcription factor identified in crustaceans and 1 of only a small number of such proteins identified in any organism undergoing diapause.