Identification of a novel gene encoding the trehalose phosphate synthase in the cotton bollworm, Helicoverpa armigera.
Trehalose and trehalose metabolism are crucial for insect development. We measured the content of polyhydric compounds in the hemolymph of diapause- and nondiapause-destined individuals of Helicoverpa armigera. We found that the trehalose content is much higher in diapause-destined individuals than that in nondiapause individuals. The activity of trehalose-6-phosphate synthase (TPS) during H. armigera larval-pupal development is significantly higher in diapause-type individuals and is closely correlated with the changes in the trehalose content. The cDNA encoding TPS, which converts uridine-5'-diphosphoglucose and glucose-6-phosphate to trehalose-6-phosphate, was cloned from the fat body of H. armigera using rapid amplification of cDNA ends (RACE). The molecular characterization of the cDNA revealed that the mRNA encodes a precursor polypeptide of 826-amino-acid residues, containing Har-TPS at residues 6-507 and a putative trehalose-6-phosphate phosphatase, which converts trehalose-6-phosphate into free trehalose, at residues 512-783. The Har-TPS precursor polypeptide shows 73% identity with that of Drosophila melanogaster. The presence of a 2.8 kb transcript in the fat body and ovary was detected with a northern blot. The Har-TPS mRNA was detected at high levels in the late stage of sixth larval instar and the early middle stage of diapause-destined pupae, which are most likely to respond the changes in TPS activity and trehalose in the hemolymph. The Har-TPS protein was successfully overexpressed in the Bombyx mori baculovirus expression system, and the catalytic activity of Har-TPS was found to be approximately 5-fold higher in B. mori blood infected by the recombined-baculovirus than the control. When diapause is broken, the trehalose content drops significantly and glucose increases rapidly. These results suggest that trehalose is involved in regulating H. armigera pupal diapause.
Xu
Jun
J
State Key Laboratory of Biocontrol and School of Life Sciences, Sun Yat-Sen (Zhongshan) University, Guangzhou 510275, China.
Bao
Bin
B
Zhang
Zhi-Fang
ZF
Yi
Yong-Zhu
YZ
Xu
Wei-Hua
WH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005216:Fat Body / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D017344:Genes, Insect; D005964:Glucosyltransferases / Q000235:genetics*; D003368:Gossypium / Q000235:genetics / Q000378:metabolism; D006458:Hemolymph / Q000201:enzymology; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008675:Metamorphosis, Biological / Q000235:genetics; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D016366:Open Reading Frames; D011679:Pupa / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013403:Sugar Phosphates / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution; D014199:Trehalose / Q000031:analogs & derivatives / Q000235:genetics / Q000378:metabolism
D018076:DNA, Complementary; D012333:RNA, Messenger; D013403:Sugar Phosphates; C082722:trehalose-6-phosphate; D014199:Trehalose; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase