[Show abstract][Hide abstract] ABSTRACT: Production of lysophosphatidic acid (LPA) is the first step in the de novo pathway for glycerolipid biosynthesis, which is mainly catalyzed by the glycerol-3-phosphate acyltransferases (GPATs; EC22.214.171.124). DHAPAT (EC126.96.36.199) also contributes in a minor way, using dihydroxyacetone phosphate as substrate. Final products and intermediates of the glycerolipid synthesis pathway are the main structural components of cellular membranes, and provide signalling molecules that regulate diverse biological processes, including cell proliferation, differentiation and growth. Here we identified the four orthologs of the mammalian GPATs (1-4) and DHAPAT in Xenopus, including a novel, short variant of GPAT2, and analyzed their expression pattern during embryonic development. Xenopus GPAT1/2 localized to mitochondria, while GPAT3/4 associated with the endoplasmic reticulum. All are similarly expressed in the early embryonic nervous system. A more tissue specific pattern emerges during organogenesis, including liver expression for GPAT1/4, and testis expression for GPAT2. All acyltransferases were expressed in kidney, though GPAT3 was excluded from the pronephric ducts. Our results suggest important roles of GPATs and DHAPAT during early organogenesis.
[Show abstract][Hide abstract] ABSTRACT: INhibitor of Growth (ING) proteins belong to a large family of plant homeodomain finger-containing proteins important in epigenetic regulation and carcinogenesis. We have previously shown that ING1 and ING2 expression is regulated by thyroid hormone (TH) during metamorphosis of the Xenopus laevis tadpole. The present study investigates the possibility that ING proteins modulate TH action.
Tadpoles expressing a Xenopus ING2 transgene (Trans(ING2)) were significantly smaller than tadpoles not expressing the transgene (Trans(GFP)). When exposed to 10 nM 3,5,3'-triiodothyronine (T(3)), premetamorphic Trans(ING2) tadpoles exhibited a greater reduction in tail, head, and brain areas, and a protrusion of the lower jaw than T(3)-treated Trans(GFP) tadpoles. Quantitative real time polymerase chain reaction (QPCR) demonstrated elevated TH receptor β (TRβ) and TH/bZIP transcript levels in Trans(ING2) tadpole tails compared to Trans(GFP) tadpoles while TRα mRNAs were unaffected. In contrast, no difference in TRα, TRβ or insulin-like growth factor (IGF2) mRNA abundance was observed in the brain between Trans(ING2) and Trans(GFP) tadpoles. All of these transcripts, except for TRα mRNA in the brain, were inducible by the hormone in both tissues. Oocyte transcription assays indicated that ING proteins enhanced TR-dependent, T(3)-induced TRβ gene promoter activity. Examination of endogenous T(3)-responsive promoters (TRβ and TH/bZIP) in the tail by chromatin immunoprecipitation assays showed that ING proteins were recruited to TRE-containing regions in T(3)-dependent and independent ways, respectively. Moreover, ING and TR proteins coimmunoprecipitated from tail protein homogenates derived from metamorphic climax animals.
We show for the first time that ING proteins modulate TH-dependent responses, thus revealing a novel role for ING proteins in hormone signaling. This has important implications for understanding hormone influenced disease states and suggests that the induction of ING proteins may facilitate TR function during metamorphosis in a tissue-specific manner.
PLoS ONE 01/2011; 6(12):e28658. · 3.53 Impact Factor