-
[show abstract]
[hide abstract]
ABSTRACT: A new insect member of the signal transducer and activator of transcription (STAT) family of transcription factors, Hyphantria cunea STAT (HcSTAT), was cloned from the lepidopteran H. cunea. The domain involved in DNA interaction and the Src homology 2 (SH2) domain were well conserved. During all developmental stages, the gene was expressed at a low level in the haemocytes, fat body cells, midgut, epidermis and Malpighian tubules. The haemocytes and Malpighian tubules showed transcriptional activation of HcSTAT upon Gram-negative and Gram-positive bacterial challenges. These challenges increased the induction and nuclear translocation of the HcSTAT protein that recognizes a STAT target site in H. cunea haemocytes. In vivo treatment with sodium orthovanadate translocated HcSTAT to the haemocyte nucleus. This study shows the involvement of the haemocyte Janus kinase/STAT pathway after microbial infection in lepidopteran insects.
Insect Molecular Biology 09/2011; 20(6):723-32. · 2.53 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A manganese superoxide dismutase (MnSOD) gene was cloned from the fall webworm, Hyphantria cunea. MnSOD cDNAs encode precursor proteins of 215 amino acid residues. H. cunea MnSOD possesses the metal binding ligands of 3 histidines and 1 aspartic acid common to MnSODs. The deduced amino acid sequences of the H. cunea MnSOD cDNA showed 76% identity to Bombyx mori MnSOD and 56-62% identity to MnSOD sequences from other species. MnSOD and copper/zinc superoxide dismutase (Cu/ZnSOD) is expressed in all tissues of H. cunea. MnSOD expression changed at a trace-level in infected larvae, while Cu/ZnSOD expression strongly changed against Gram-positive and Gram-negative bacteria, and fungi. Environmental stresses such as different artificial photoperiods (24L:0D), ultraviolet irradiation (312 nm), and starvation condition increased Cu/ZnSOD expression, MnSOD expression, on the other hand, was increased by starvation. Moreover, MnSOD and Cu/ZnSOD expression showed no significant change in the 0L:24D condition. MnSOD and Cu/ZnSOD expression in H. cunea also significantly increased at high (37°C) and low (4°C) temperature. Oxidative stress induced by 10% H(2)O(2) reduced the expression levels of MnSOD and Cu/ZnSOD. However, paraquat-induced oxidative stress reduced MnSOD expression but increased Cu/ZnSOD expression. These results suggest that Cu/ZnSOD may play a larger role than MnSOD as a superoxide anion scavenger against oxidative stress in H. cunea.
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 12/2010; 157(4):343-50. · 1.61 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A full-length clone corresponding to attacin was isolated from a cDNA library made from fat body of immunized Hyphantria cunea larvae. This newly isolated attacin B shows characteristics different from those previously reported for attacin A. The two attacin cDNAs encode precursor proteins of 233 and 248 amino acid residues, respectively. The two attacins show 45.9% identity at the amino acid level, and 35.2% identity at the nucleotide level. Attacins A and B of H. cunea show significant identities with the attacins of Lepidoptera. Attacin B is a typical glycine-rich protein, while attacin A is leucine-rich. Attacin B is expressed from last instar larvae to adult, while attacin A showed stage-specific expression during the prepupal and pupal stages. Attacins A and B are predicted to have different secondary structure in that attacin A has no tendency to form helices but attacin B contains a substantial number of helices. Attacin A is induced at a trace level in infected larvae, while attacin B is strongly induced against Gram-positive and negative bacteria, fungi, and viruses. The attacin B transcripts were detected in fat body, epidermis and hemocytes after injection with Escherichia coli, Citrobacter freundii, or Candida albicans, but not in the midgut and Malpighian tubule. Recombinant attacin A showed no antibacterial activity, while recombinant attacin B showed strong antibacterial activity in proportion to the amount of the protein injected.
Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology 11/2008; 151(2):213-20. · 1.92 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: cDNA clones for two of the yolk proteins, YP1 and YP2, produced by the fat body of the moth, Hyphantria cunea, were sequenced and found to be homologous to the follicular epithelium yolk proteins of pyralid moths. Both cDNA clones coded for polypeptides of 290 residues and the deduced amino acid sequence identity between YP1 and YP2 was very high (79.0%). Analysis of the secondary structure of the predicted polypeptides suggests that YP1 and YP2 do not form heteromeric proteins because of differences in secondary structure due to the lack of alpha helices in YP1. Northern blot analysis showed that the transcripts for YP1 (1.2 kb) and YP2 (1.1 kb) were present primarily in the female fat body with only trace levels detectable in the ovary of the adult female. In a developmental study, the YP1 and YP2 transcripts were first detectable in 10-day-old pupae and increased into the adult stage. These results suggest that the YP1 and YP2 genes in H. cunea have been recruited to replace the vitellogenin gene as the primary source of yolk proteins. During this process they have acquired a modified pattern of expression that is different from homologous genes reported in pyralid moths. The assessment of the evolution of proteinaceous yolk in these moths should serve as an excellent model for the evolution of gene recruitment.
Insect Molecular Biology 09/2003; 12(4):383-92. · 2.53 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Storage protein-1 (HcSP-1) is a major storage protein found in the hemolymph and fat body of Hyphantria cunea. HcSP-1 has a high methionine (6.0%) and low aromatic amino acid content (8.5%) (Cheon et al., 1998). In this study, the accumulation and expression of HcSP-1 in ovary was investigated using biochemical and immunocytochemical methods. HcSP-1 was detected in the ovaries in 6-day-old pupae and accumulated toward the end of pupal life, when HcSP-1 transcripts were detectable by Northern blot analysis and RT-PCR. In situ hybridization showed that the HcSP-1 mRNA was located in the nurse cells and follicular epithelial cells, but not in the oocyte. Though most of the HcSP-1 that is incorporated in the yolk bodies of the oocyte is probably sequestered from the surrounding hemolymph, HcSP-1 is an important yolk protein contributing to early yolk body formation before the development of patency by the follicular epithelium.
Archives of Insect Biochemistry and Physiology 12/2001; 48(3):111-20. · 1.36 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We isolated and sequenced a cDNA clone corresponding to storage protein-2 (SP-2) from the fall webworm, Hyphantria cunea. The cDNA for SP-2 (2572 bp) codes for a 747-residue protein with a predicted molecular mass of 88.5 kDa. The calculated isoelectric point is 7.6. Multiple alignment analysis of amino acid sequence revealed that SP-2 is most similar to BJHSP2 (74.3% identity). According to both the phylogenetic analyses and criteria for amino acid composition, SP-2 belongs to the subfamily of moderately methionine-rich storage proteins (3.2% methionine, 11.8% aromatic amino acid). Topical application of the JH analog, methoprene, after head ligation of larvae, suppressed transcription of the SP-2 gene, indicating hormonal effects at the transcriptional level. The SP-2 transcript was detected by Northern blot analysis in Malpighian tubules, in addition to the fat body where it was most abundant. The local expression of SP-2 in Malpighian tubules suggests that it may have some function in that organ.
Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology 06/2001; 129(1):97-107. · 1.92 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A full-length clone corresponding to attacin was isolated from a cDNA library made from fat body of immunized Hyphantria cunea larvae. This newly isolated attacin B shows characteristics different from those previously reported for attacin A. The two attacin cDNAs encode precursor proteins of 233 and 248 amino acid residues, respectively. The two attacins show 45.9% identity at the amino acid level, and 35.2% identity at the nucleotide level. Attacins A and B of H. cunea show significant identities with the attacins of Lepidoptera. Attacin B is a typical glycine-rich protein, while attacin A is leucine-rich. Attacin B is expressed from last instar larvae to adult, while attacin A showed stage-specific expression during the prepupal and pupal stages. Attacins A and B are predicted to have different secondary structure in that attacin A has no tendency to form helices but attacin B contains a substantial number of helices. Attacin A is induced at a trace level in infected larvae, while attacin B is strongly induced against Gram-positive and negative bacteria, fungi, and viruses. The attacin B transcripts were detected in fat body, epidermis and hemocytes after injection with Escherichia coli, Citrobacter freundii, or Candida albicans, but not in the midgut and Malpighian tubule. Recombinant attacin A showed no antibacterial activity, while recombinant attacin B showed strong antibacterial activity in proportion to the amount of the protein injected.
Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology.
-
-
[show abstract]
[hide abstract]
ABSTRACT: A manganese superoxide dismutase (MnSOD) gene was cloned from the fall webworm, Hyphantria cunea. MnSOD cDNAs encode precursor proteins of 215 amino acid residues. H. cunea MnSOD possesses the metal binding ligands of 3 histidines and 1 aspartic acid common to MnSODs. The deduced amino acid sequences of the H. cunea MnSOD cDNA showed 76% identity to Bombyx mori MnSOD and 56–62% identity to MnSOD sequences from other species. MnSOD and copper/zinc superoxide dismutase (Cu/ZnSOD) is expressed in all tissues of H. cunea. MnSOD expression changed at a trace-level in infected larvae, while Cu/ZnSOD expression strongly changed against Gram-positive and Gram-negative bacteria, and fungi. Environmental stresses such as different artificial photoperiods (24L:0D), ultraviolet irradiation (312 nm), and starvation condition increased Cu/ZnSOD expression, MnSOD expression, on the other hand, was increased by starvation. Moreover, MnSOD and Cu/ZnSOD expression showed no significant change in the 0L:24D condition. MnSOD and Cu/ZnSOD expression in H. cunea also significantly increased at high (37 °C) and low (4 °C) temperature. Oxidative stress induced by 10% H2O2 reduced the expression levels of MnSOD and Cu/ZnSOD. However, paraquat-induced oxidative stress reduced MnSOD expression but increased Cu/ZnSOD expression. These results suggest that Cu/ZnSOD may play a larger role than MnSOD as a superoxide anion scavenger against oxidative stress in H. cunea.
Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology.
