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A eukaryotic initiation factor 5C is upregulated during metamorphosis in the cotton bollworm, Helicoverpa armigera.

Dong DJ, Wang JX, Zhao XF - BMC Dev. Biol. (2009)

Bottom Line: Ha-eIF5C expression was upregulated by 20-hydroxyecdysone (20E).Furthermore, the transcription of Ha-eIF5C was down regulated after silencing of ecdysteroid receptor (EcR) or Ultraspiracle protein (USP) by RNAi.These results suggested that during metamorphosis of the cotton bollworm, Ha-eIF5C was upregulated by 20E through the EcR and USP transcription factors.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of Ministry of Education, School of Life Sciences, Shandong University, Jinan 250100, Shandong, PR China. dujuandong@yahoo.com.cn

ABSTRACT

Background: The orthologs of eukaryotic initiation factor 5C (eIF5C) are essential to the initiation of protein translation, and their regulation during development is not well known.

Results: A cDNA encoding a polypeptide of 419 amino acids containing an N-terminal leucine zipper motif and a C-terminal eIF5C domain was cloned from metamorphic larvae of Helicoverpa armigera. It was subsequently named Ha-eIF5C. Quantitative real-time PCR (QRT-PCR) revealed a high expression of the mRNA of Ha-eIF5C in the head-thorax, integument, midgut, and fat body during metamorphosis. Immunohistochemistry suggested that Ha-eIF5C was distributed into both the cytoplasm and the nucleus in the midgut, fat body and integument. Ha-eIF5C expression was upregulated by 20-hydroxyecdysone (20E). Furthermore, the transcription of Ha-eIF5C was down regulated after silencing of ecdysteroid receptor (EcR) or Ultraspiracle protein (USP) by RNAi.

Conclusion: These results suggested that during metamorphosis of the cotton bollworm, Ha-eIF5C was upregulated by 20E through the EcR and USP transcription factors.

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Complete cDNA sequence and deduced amino acid sequence of Ha-eIF5C. The underlined amino acid sequences indicate predicted phosphorylation sites. Protein kinase C phosphorylation sites (2–4; 17–19; 91–93; 107–109; 193–195; 335–337; 387–389); tyrosine kinase phosphorylation sites (18–26; 345–351); casein kinase II phosphorylation sites (58–61; 100–103; 303–306; 407–410; 414–417). The putative N-glycosylation site is shaded. Predicted N-myristoylation sites are in block.
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Figure 1: Complete cDNA sequence and deduced amino acid sequence of Ha-eIF5C. The underlined amino acid sequences indicate predicted phosphorylation sites. Protein kinase C phosphorylation sites (2–4; 17–19; 91–93; 107–109; 193–195; 335–337; 387–389); tyrosine kinase phosphorylation sites (18–26; 345–351); casein kinase II phosphorylation sites (58–61; 100–103; 303–306; 407–410; 414–417). The putative N-glycosylation site is shaded. Predicted N-myristoylation sites are in block.

Mentions: Based on the fragment of Ha-eIF5C obtained from suppression subtractive hybridization (SSH), the 5' end fragment was obtained using specific reverse primer eIF5CR and the T3 primer. The 3' end fragment was amplified with the specific primer eIF5CF and the T7 primer. The full-length eIF5C of H. armegera (1675 bp) was obtained through an assemblage of overlapping nucleic acids. This included a 57 bp 5' untranslated region (UTR), a 1260 bp open reading frame and a 340 bp untranslated region in the 3' UTR, with a 18 bp poly A tail. The ORF encoded a 419 amino acid protein with a calculated molecular mass of 48 kDa and a predicated isoelectric point of 6.05. Moreover, there were some putative post-translational modification sites including seven protein kinase c phosphorylation sites, two tyrosine kinase phosphorylation sites, three N-myristoylation sites, five casein kinase II phosphorylation sites and one N-glycosylation site (Fig. 1).


A eukaryotic initiation factor 5C is upregulated during metamorphosis in the cotton bollworm, Helicoverpa armigera.

Dong DJ, Wang JX, Zhao XF - BMC Dev. Biol. (2009)

Complete cDNA sequence and deduced amino acid sequence of Ha-eIF5C. The underlined amino acid sequences indicate predicted phosphorylation sites. Protein kinase C phosphorylation sites (2–4; 17–19; 91–93; 107–109; 193–195; 335–337; 387–389); tyrosine kinase phosphorylation sites (18–26; 345–351); casein kinase II phosphorylation sites (58–61; 100–103; 303–306; 407–410; 414–417). The putative N-glycosylation site is shaded. Predicted N-myristoylation sites are in block.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2667495&req=5

Figure 1: Complete cDNA sequence and deduced amino acid sequence of Ha-eIF5C. The underlined amino acid sequences indicate predicted phosphorylation sites. Protein kinase C phosphorylation sites (2–4; 17–19; 91–93; 107–109; 193–195; 335–337; 387–389); tyrosine kinase phosphorylation sites (18–26; 345–351); casein kinase II phosphorylation sites (58–61; 100–103; 303–306; 407–410; 414–417). The putative N-glycosylation site is shaded. Predicted N-myristoylation sites are in block.
Mentions: Based on the fragment of Ha-eIF5C obtained from suppression subtractive hybridization (SSH), the 5' end fragment was obtained using specific reverse primer eIF5CR and the T3 primer. The 3' end fragment was amplified with the specific primer eIF5CF and the T7 primer. The full-length eIF5C of H. armegera (1675 bp) was obtained through an assemblage of overlapping nucleic acids. This included a 57 bp 5' untranslated region (UTR), a 1260 bp open reading frame and a 340 bp untranslated region in the 3' UTR, with a 18 bp poly A tail. The ORF encoded a 419 amino acid protein with a calculated molecular mass of 48 kDa and a predicated isoelectric point of 6.05. Moreover, there were some putative post-translational modification sites including seven protein kinase c phosphorylation sites, two tyrosine kinase phosphorylation sites, three N-myristoylation sites, five casein kinase II phosphorylation sites and one N-glycosylation site (Fig. 1).

Bottom Line: Ha-eIF5C expression was upregulated by 20-hydroxyecdysone (20E).Furthermore, the transcription of Ha-eIF5C was down regulated after silencing of ecdysteroid receptor (EcR) or Ultraspiracle protein (USP) by RNAi.These results suggested that during metamorphosis of the cotton bollworm, Ha-eIF5C was upregulated by 20E through the EcR and USP transcription factors.

View Article: PubMed Central - HTML - PubMed

Affiliation: The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of Ministry of Education, School of Life Sciences, Shandong University, Jinan 250100, Shandong, PR China. dujuandong@yahoo.com.cn

ABSTRACT

Background: The orthologs of eukaryotic initiation factor 5C (eIF5C) are essential to the initiation of protein translation, and their regulation during development is not well known.

Results: A cDNA encoding a polypeptide of 419 amino acids containing an N-terminal leucine zipper motif and a C-terminal eIF5C domain was cloned from metamorphic larvae of Helicoverpa armigera. It was subsequently named Ha-eIF5C. Quantitative real-time PCR (QRT-PCR) revealed a high expression of the mRNA of Ha-eIF5C in the head-thorax, integument, midgut, and fat body during metamorphosis. Immunohistochemistry suggested that Ha-eIF5C was distributed into both the cytoplasm and the nucleus in the midgut, fat body and integument. Ha-eIF5C expression was upregulated by 20-hydroxyecdysone (20E). Furthermore, the transcription of Ha-eIF5C was down regulated after silencing of ecdysteroid receptor (EcR) or Ultraspiracle protein (USP) by RNAi.

Conclusion: These results suggested that during metamorphosis of the cotton bollworm, Ha-eIF5C was upregulated by 20E through the EcR and USP transcription factors.

Show MeSH