New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm
Abstract Background Exploring whether and how mutation of silk protein contributes to subsequent re-allocation of nitrogen, and impacts on the timing of silk gland degradation, is important to understand silk gland biology. Rapid development and wide application of genome editing approach in the sil...
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doaj-31c614e9b10b49e3b8e9fce59ea52ddd2020-11-25T00:49:17ZengBMCBMC Genomics1471-21642018-03-0119111010.1186/s12864-018-4602-4New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkwormYong Cui0Yanan Zhu1Yongjian Lin2Lei Chen3Qili Feng4Wen Wang5Hui Xiang6Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal UniversityGuangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal UniversityGuangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal UniversityCenter for Ecological and Environmental Sciences, Northwestern Polytechnical UniversityGuangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal UniversityCenter for Ecological and Environmental Sciences, Northwestern Polytechnical UniversityGuangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal UniversityAbstract Background Exploring whether and how mutation of silk protein contributes to subsequent re-allocation of nitrogen, and impacts on the timing of silk gland degradation, is important to understand silk gland biology. Rapid development and wide application of genome editing approach in the silkworm provide us an opportunity to address these issues. Results Using CRISPR/Cas9 system, we successfully performed genome editing of Bmfib-H. The loss-of-function mutations caused naked pupa and thin cocoon mutant phenotypes. Compared with the wild type, the posterior silk gland of mutant showed obviously degraded into fragments in advance of programmed cell death of silk gland cells. Comparative transcriptomic analyses of silk gland at the fourth day of the fifth instar larval stage(L5D4)identified 1456 differential expressed genes (DEGs) between posterior silk gland (PSG) and mid silk gland (MSG) and 1388 DEGs between the mutant and the wild type. Hierarchical clustering of all the DEGs indicated a remarkable down-regulated and an up-regulated gene clade in the mutant silk glands, respectively. Down-regulated genes were overrepresented in the pathways involved in cancer, DNA replication and cell proliferation. Intriguingly, up-regulated DEGs are significantly enriched in the proteasome. By further comparison on the transcriptome of MSG and PSG between the wild type and the mutant, we consistently observed that up-regulated DEGs in the mutant PSG were enriched in protein degrading activity and proteasome. Meantime, we observed a series of up-regulated genes involved in autophagy. Since these protein degradation processes would be normally occur after the spinning time, the results suggesting that these progresses were activated remarkably ahead of schedule in the mutant. Conclusions Accumulation of abnormal fib-H protein might arouse the activation of proteasomes as well as autophagy process, to promote the rapid degradation of such abnormal proteins and the silk gland cells. Our study therefore proposes a subsequent process of protein and partial cellular degradation caused by mutation of silk protein, which might be helpful for understanding its impact of the silk gland biological process, and further exploration the re-allocation of nitrogen in the silkworm.http://link.springer.com/article/10.1186/s12864-018-4602-4Bombyx moriCRISPR/Cas9TranscriptomeBmfib-HProteasomeAutophagy |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yong Cui Yanan Zhu Yongjian Lin Lei Chen Qili Feng Wen Wang Hui Xiang |
spellingShingle |
Yong Cui Yanan Zhu Yongjian Lin Lei Chen Qili Feng Wen Wang Hui Xiang New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm BMC Genomics Bombyx mori CRISPR/Cas9 Transcriptome Bmfib-H Proteasome Autophagy |
author_facet |
Yong Cui Yanan Zhu Yongjian Lin Lei Chen Qili Feng Wen Wang Hui Xiang |
author_sort |
Yong Cui |
title |
New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm |
title_short |
New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm |
title_full |
New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm |
title_fullStr |
New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm |
title_full_unstemmed |
New insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm |
title_sort |
new insight into the mechanism underlying the silk gland biological process by knocking out fibroin heavy chain in the silkworm |
publisher |
BMC |
series |
BMC Genomics |
issn |
1471-2164 |
publishDate |
2018-03-01 |
description |
Abstract Background Exploring whether and how mutation of silk protein contributes to subsequent re-allocation of nitrogen, and impacts on the timing of silk gland degradation, is important to understand silk gland biology. Rapid development and wide application of genome editing approach in the silkworm provide us an opportunity to address these issues. Results Using CRISPR/Cas9 system, we successfully performed genome editing of Bmfib-H. The loss-of-function mutations caused naked pupa and thin cocoon mutant phenotypes. Compared with the wild type, the posterior silk gland of mutant showed obviously degraded into fragments in advance of programmed cell death of silk gland cells. Comparative transcriptomic analyses of silk gland at the fourth day of the fifth instar larval stage(L5D4)identified 1456 differential expressed genes (DEGs) between posterior silk gland (PSG) and mid silk gland (MSG) and 1388 DEGs between the mutant and the wild type. Hierarchical clustering of all the DEGs indicated a remarkable down-regulated and an up-regulated gene clade in the mutant silk glands, respectively. Down-regulated genes were overrepresented in the pathways involved in cancer, DNA replication and cell proliferation. Intriguingly, up-regulated DEGs are significantly enriched in the proteasome. By further comparison on the transcriptome of MSG and PSG between the wild type and the mutant, we consistently observed that up-regulated DEGs in the mutant PSG were enriched in protein degrading activity and proteasome. Meantime, we observed a series of up-regulated genes involved in autophagy. Since these protein degradation processes would be normally occur after the spinning time, the results suggesting that these progresses were activated remarkably ahead of schedule in the mutant. Conclusions Accumulation of abnormal fib-H protein might arouse the activation of proteasomes as well as autophagy process, to promote the rapid degradation of such abnormal proteins and the silk gland cells. Our study therefore proposes a subsequent process of protein and partial cellular degradation caused by mutation of silk protein, which might be helpful for understanding its impact of the silk gland biological process, and further exploration the re-allocation of nitrogen in the silkworm. |
topic |
Bombyx mori CRISPR/Cas9 Transcriptome Bmfib-H Proteasome Autophagy |
url |
http://link.springer.com/article/10.1186/s12864-018-4602-4 |
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