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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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QRT-PCR analysis of gene expression in five tissues during larval feeding, molting and metamorphosis. ht: head-thorax, int: integument, mg: midgut, fb: fat body, hc: haemocytes. 5th24 = 5th instar larvae 24 h after ecdysis, 5th36 = 5th instar larvae 36 h after ecdysis (5th-HCS, with head capsule slippage), 6th72 = 72 h after ecdysis (wanderting 0 d, metamorphically committed larvae). Error bars represent the standard deviation in three replicates. An asterisk indicates significant differences (Student' s t-test, *: p < 0.05).
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Figure 4: QRT-PCR analysis of gene expression in five tissues during larval feeding, molting and metamorphosis. ht: head-thorax, int: integument, mg: midgut, fb: fat body, hc: haemocytes. 5th24 = 5th instar larvae 24 h after ecdysis, 5th36 = 5th instar larvae 36 h after ecdysis (5th-HCS, with head capsule slippage), 6th72 = 72 h after ecdysis (wanderting 0 d, metamorphically committed larvae). Error bars represent the standard deviation in three replicates. An asterisk indicates significant differences (Student' s t-test, *: p < 0.05).

Mentions: To study the tissue distribution of Ha-eIF5C, the total RNA of the head-thorax, integument, midgut, fatbody and haemocyte were extracted from 5th 24 h (5th instar larvae 24 h after ecdysis), 5th-HCS (5th instar larvae 36 h after ecdysis, with head capsule slippage, HCS) and 6th 72 h (72 h after ecdysis, wandering 0 d, metamorphically committed larva) stage. As shown in Fig. 4, Ha-eIF5C transcript was detected at a high level in the head-thorax, integument, midgut and fat body but not in haemocytes in metamorphosis stage. QRT-PCR was utilized to analyze the expression of Ha-eIF5C in developmental midgut and fat body. The results showed that there was an obvious increase in the level of Ha-eIF5C transcript during metamorphosis. The immunoblotting revealed that the expression of Ha-eIF5C protein agreed with the mRNA transcription (Fig. 5).


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)

QRT-PCR analysis of gene expression in five tissues during larval feeding, molting and metamorphosis. ht: head-thorax, int: integument, mg: midgut, fb: fat body, hc: haemocytes. 5th24 = 5th instar larvae 24 h after ecdysis, 5th36 = 5th instar larvae 36 h after ecdysis (5th-HCS, with head capsule slippage), 6th72 = 72 h after ecdysis (wanderting 0 d, metamorphically committed larvae). Error bars represent the standard deviation in three replicates. An asterisk indicates significant differences (Student' s t-test, *: p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: QRT-PCR analysis of gene expression in five tissues during larval feeding, molting and metamorphosis. ht: head-thorax, int: integument, mg: midgut, fb: fat body, hc: haemocytes. 5th24 = 5th instar larvae 24 h after ecdysis, 5th36 = 5th instar larvae 36 h after ecdysis (5th-HCS, with head capsule slippage), 6th72 = 72 h after ecdysis (wanderting 0 d, metamorphically committed larvae). Error bars represent the standard deviation in three replicates. An asterisk indicates significant differences (Student' s t-test, *: p < 0.05).
Mentions: To study the tissue distribution of Ha-eIF5C, the total RNA of the head-thorax, integument, midgut, fatbody and haemocyte were extracted from 5th 24 h (5th instar larvae 24 h after ecdysis), 5th-HCS (5th instar larvae 36 h after ecdysis, with head capsule slippage, HCS) and 6th 72 h (72 h after ecdysis, wandering 0 d, metamorphically committed larva) stage. As shown in Fig. 4, Ha-eIF5C transcript was detected at a high level in the head-thorax, integument, midgut and fat body but not in haemocytes in metamorphosis stage. QRT-PCR was utilized to analyze the expression of Ha-eIF5C in developmental midgut and fat body. The results showed that there was an obvious increase in the level of Ha-eIF5C transcript during metamorphosis. The immunoblotting revealed that the expression of Ha-eIF5C protein agreed with the mRNA transcription (Fig. 5).

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