Role of Trehalose Phosphate Synthase in Anoxia Tolerance and Development in Drosophila melanogaster

Chen, Qiaofang; Ma, Enbo; Behar, Kevin L.; Xu, Tian; Haddad, Gabriel G.

doi:10.1074/jbc.m109479200

Cited by 162 publications

(144 citation statements)

References 24 publications

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“…This central role of T6P may be more widely conserved than previously imagined. Beyond yeasts and plants, it could well be conserved in insects: Drosophila mutated in the TPS has recently been shown to be embryo lethal (48). There exists a particular pressing need to find the site and details of the T6P interaction in all these organisms.…”

Section: T6p Levels Determine the Accumulation Of Respiratory Intermementioning

confidence: 99%

Trehalose 6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana

Schluepmann

Pellny²,

Dijken³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

446

497

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Section: T6p Levels Determine the Accumulation Of Respiratory Intermementioning

confidence: 99%

Trehalose 6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana

Schluepmann

Pellny²,

Dijken³

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

446

497

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“…Even in so-called "normal" organisms, trehalose synthesis is elevated under stress conditions (most recently reviewed in Ref. 1), including dehydration, stationary phase culture, salt-stress (5), heat-shock (2), high pressure (6), extreme cold (7), exposure to oxygen radicals (8,9), and anoxia (10).…”

mentioning

confidence: 99%

The Donor Subsite of Trehalose-6-phosphate Synthase

Gibson

Tarling

Roberts

et al. 2004

Journal of Biological Chemistry

106

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Trehalose is an unusual non-reducing disaccharide that plays a variety of biological roles, from food storage to cellular protection from environmental stresses such as desiccation, pressure, heat-shock, extreme cold, and oxygen radicals. It is also an integral component of the cell-wall glycolipids of mycobacteria. The primary enzymatic route to trehalose first involves the transfer of glucose from a UDP-glucose donor to glucose-6-phosphate to form ␣,␣-1,1 trehalose-6-phosphate. This reaction, in which the configurations of two glycosidic bonds are set simultaneously, is catalyzed by the glycosyltransferase trehalose-6-phosphate synthase (OtsA), which acts with retention of the anomeric configuration of the UDP-sugar donor. The classification of activated sugar-dependent glycosyltransferases into approximately 70 distinct families based upon amino acid sequence similarities places OtsA in glycosyltransferase family 20 (see afmb.cnrs-mrs.fr/CAZY/). The recent 2.4 Å structure of Escherichia coli OtsA revealed a two-domain enzyme with catalysis occurring at the interface of the twin ␤/␣/␤ domains. Here we present the 2.0 Å structures of the E. coli OtsA in complex with either UDP-Glc or the non-transferable analogue UDP-2-deoxy-2-fluoroglucose. Both complexes unveil the donor subsite interactions, confirming a strong similarity to glycogen phosphorylases, and reveal substantial conformational differences to the previously reported complex with UDP and glucose 6-phosphate. Both the relative orientation of the two domains and substantial (up to 10 Å) movements of an N-terminal loop (residues 9 -22) characterize the more open "relaxed" conformation of the binary UDP-sugar complexes reported here.

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“…Like Actn null mutants, null mutants of Tps1 typically die as first instars, 30 and once again, we observed that the RNAi phenotype was similar to that of a null mutation, but with reduced penetrance.…”

Section: Resultsmentioning

confidence: 73%

Biolistics for high-throughput transformation and RNA interference inDrosophila melanogaster

Yuen

Read²,

Brubacher³

et al. 2008

Fly

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With twelve Drosophila genomes now sequenced, there is a growing need to develop higher-throughput methods for identifying the functions of the many newly identified genes. Genetic transformation and RNA interference are two technologies that have been used extensively to facilitate gene-function studies in Drosophila melanogaster, to introduce genes or block the expression of endogenous genes, respectively. Both of these technologies typically require the delivery of nucleic acids into developing insect embryos, and virtually all studies to date have relied on microinjection as the DNA delivery method of choice. In this study, we describe the use of biolistics as a higher-throughput method of nucleic acid delivery. By bombarding dechorionated D. melanogaster embryos with 1 μm gold beads coated with P-element or piggyBac transformation vectors, we observed transformation frequencies (3-4%) that are comparable to those achieved using microinjection methods, but in only a fraction of the time required for the DNA delivery. Biolistic delivery of double-stranded RNA (dsRNA) specific to a β-glucuronidase (gus) transgene resulted in a significant (71%) reduction in gus transcripts in embryos and the RNA interference (RNAi) persisted through two successive larval molts, albeit at reduced levels. DsRNAs specific to four essential genes were delivered to embryos and resulted in arrested development and phenotypes that closely match that of null mutations. These results suggest that biolistic delivery of dsRNA into embryos could be adapted for high throughput RNAi screens of early Drosophila developmental genes.

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Role of Trehalose Phosphate Synthase in Anoxia Tolerance and Development in Drosophila melanogaster

Cited by 162 publications

References 24 publications

Trehalose 6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana

Trehalose 6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana

The Donor Subsite of Trehalose-6-phosphate Synthase

Biolistics for high-throughput transformation and RNA interference inDrosophila melanogaster

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