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RISS 인기검색어
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- 저자 : Huang Qunxia (검색결과 2 건)
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m6A writer gene fl(2)d regulates the metamorphosis of silkworm Bombyx mori
Huang Qunxia,Huo Chunyue,Qian Yuxin,Zhao Chenyue,Cai Zhiyan,Zhang Kun,Zhao Xuhui,Guo Shigang,Kan Yunchao,Li Dandan 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.4
The N6-methyladenosine (m6A) is one of the most abundant posttranscriptional modifications on RNAs, which is mediated by its effector proteins—writers, readers, and erasers. Female-Lethal(2)D (Fl(2)d) is one of the key writers to stabilize the interaction between another two writers Methyltransferase-like 3 (METTL3) and Methyltransferase-like 14 (METTL14). Here with dsRNA transfection, fl(2)d was knocked down in the BmN4 cell lines of silkworm, the expression of 61 genes were significantly increased, and 86 genes were significantly decreased after fl(2)d repression, which encoded proteins related to catalytic, hydrolase activity or in metabolism of nucleic acid. To elucidate the functional role of fl(2)d during silkworm development, dsRNA of which was injected into the hemolymph of the 2-d-old fourth instar larvae, the mounting process of silkworm was delayed, the expression level of genes in metabolism, RNA processing and ecdysone signaling pathway decreased significantly after fl(2)d knockdown, such as genes encoded Ubiquitin protein ligase E3C, the nuclear pore complex protein Nup98-Nup96, and the m6A reader YTHDF3, as well as Broad Complex isoform 2 (BRC-Z2), Fushi Tarazu Factor-1 (FTZ-F1), lactate dehydrogenase (LDH) and the ecdysone-induced protein 74EF isoform A (E74A). Total m6A level of RNAs also decreased after fl(2)d repression. These indicated that decreased level of fl (2)d and YTHDF3 decay the m6A level and translation of their target genes, which might lead to the retardation of metamorphosis, and resulted in the delayed mounting of silkworm.
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Zhao Chenyue,Huang Qunxia,Qian Yuxin,Zhao Xuhui,Guo Shigang,Kan Yunchao,Li Dandan 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.3
The cuticle supports the insect body and serves as a protective barrier against pathogens and pesticides. We found that the gene CPH19, encoding the hypothetical cuticular protein 19 (CPH19) of the silkworm Bombyx mori, was highly expressed in the cuticle during larval molt. Injection of CPH19 double-stranded RNA (dsRNA) into the hemolymph of 2-day-old third instar larvae (2dLa3) accelerated insect molting at the La3-La4 and La4-La5 transitions. Nevertheless, the larvae injected with CPH19 dsRNA had difficulty shedding their cuticle and suffered from molt defects, and their exuviae were larger and heavier than those of the control group. Morphological comparison of the exuviae between CPH19 dsRNA-injected and control larvae showed that the exocuticle-endocuticle layer was thicker in the former. The effect of CPH19 dsRNA injection on gene expression was further investigated by quantitative real-time polymerase-chain reaction (qRT-PCR). Surprisingly, the mRNA levels of CPH19 and other CP genes such as CPH18, CPH20, CPR69, CPR96, CPR97, CPR98 and CPR100, as well as chitinase genes such as chs-2 and chsA, were significantly increased in the CPH19 dsRNA-injected insects compared to the control. Bioinformatic predictions and luciferase assay analyses identified a proximal promoter near the transcription start site of CPH19 with strong transcriptional activity that may be involved in an RNAbased promoter targeting mechanism mediated by small activating RNAs (saRNAs). Treatment with the insec ticide decamethrin revealed that CPH19 dsRNA-injected larvae were more resistant to insecticides than control larvae, which could be exploited for the development of new pest control strategies.
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