The Cytophaga hutchinsonii ChTPSP: First Characterized Bifunctional TPS–TPP Protein as Putative Ancestor of All Eukaryotic Trehalose Biosynthesis Proteins

Abstract

The most widely distributed pathway to synthesize trehalose in nature consists of two consecutive enzymatic reactions with a trehalose-6-P (T6P)-synthase (TPS) enzyme, producing the intermediate T6P, and a T6P-phosphatase (TPP) enzyme, which dephosphorylates T6P to produce trehalose and inorganic phosphate. In plants, these enzymes are called Class I and Class II proteins, respectively, with some Class I proteins being active enzymes. The Class II proteins possess both TPS and TPP consensus regions but appear to have lost enzymatic activity during evolution. Plants also contain an extra group of enzymes of small protein size, of which some members have been characterized as functional TPPs. These Class III proteins have less sequence similarity with the Class I and Class II proteins. Here, we characterize for the first time, by using biochemical analysis and yeast growth complementation assays, the existence of a natural TPS-TPP bifunctional enzyme found in the bacterial species Cytophaga hutchinsonii. Through phylogenetic analysis, we show that prokaryotic genes such as ChTPSP might be the ancestor of the eukaryotic trehalose biosynthesis genes. Second, we show that plants have recruited during evolution, possibly by horizontal transfer from bacteria such as Rhodoferax ferrireducens, a new type of small protein, encoding TPP activity, which have been named Class III proteins. RfTPP has very high TPP activity upon expression in yeast. Finally, we demonstrate that TPS gene duplication, the recruitment of the Class III enzymes, and recruitment of an N-terminal regulatory element, which regulates the Class I enzyme activity in higher plants, were initiated very early in eukaryan evolution as the three classes of trehalose biosynthesis genes are already present in the alga Ostreococcus tauri.

The Class II Trehalose 6-phosphate Synthase Gene PvTPS9 Modulates Trehalose Metabolism in Phaseolus vulgaris Nodules

“…One subgroup contained only PvTPS3 , its sequence being closer to TPS genes from other legume species such as Lotus japonicus ; the other subgroup contained PvTPS1 and PvTPS2 (Figure 1A). As found in other plants, common bean Class II TPS proteins were clustered in subfamilies containing members from both monocots and dicots (Figure 1A), suggesting that the Class II TPS of common bean arose from ancient gene duplication events followed by specialization, as has been proposed for Arabidopsis thaliana Class II TPS (Ramon et al, 2009; Avonce et al, 2010). In silico analysis of the P. vulgaris transcriptome (Ramírez et al, 2005; Hernández et al, 2007; DFCI, http://compbio.dfci.harvard.edu/tgi/) revealed that all TPS and TPP genes in common bean plants were expressed, either during some part of plant development or in a tissue-specific manner.…”

“…In plants, multi-gene families encode TPS and TPP (Leyman et al, 2001). TPS genes include Class I genes, which usually encode catalytically active TPS enzymes, and Class II genes, which encode proteins that do not possess TPS or TPP enzymatic activity but contain TPS and TPP domains (Avonce et al, 2010). The Class III family encodes functional TPP enzymes (Vogel et al, 1998; Vandesteene et al, 2012).…”

A Tale of Two Sugars: Trehalose 6-Phosphate and Sucrose

“…TPS and TPP genes have now been identified in species from all major plant taxa, indicating that trehalose metabolism is probably universal in the plant kingdom (Avonce et al, 2006(Avonce et al, , 2010Lunn, 2007;Lunn et al, 2014). Arabidopsis has 11 TPS (AtTPS1-AtTPS11) and 10 TPP (AtTPPA-AtTPPJ) genes (Arabidopsis Genome Initiative, 2000; Leyman et al, 2001).…”

Up-regulation of biosynthetic processes associated with growth by trehalose 6-phosphate

“…A new distinct, previously unknown function of the pathway appears to have developed early in plant evolution. 3 Clues to this function have been provided in mutant and transgenic plants, which show an indispensable role for the pathway in the utilization of carbon and in growth and development. 4 Until recently a mechanistic basis to explain this function was not known.…”