|
|
|
|
|
| Amplification and function analysis of N6-adenine-specific DNA methyltransferase gene in Nilaparvata lugens |
| ZHANG Jiao, XING Yan-ru, HOU Bo-feng, YUAN Zhu-ting, LI Yao, JIE Wen-cai, SUN Yang, LI Fei |
1、Department of Entomology, College of Plant Protection, Nanjing Agricultural University/Key Laboratory of Monitoring and
Management of Plant Diseases and Insects, Ministry of Agriculture, Nanjing 210095, P.R.China
2、Key Lab of Agricultural Entomology, Ministry of Agriculture/Institute of Insect Sciences, Zhejiang University, Hangzhou 310058,
P.R.China |
|
|
|
摘要 Methylation of the N6 position of adenine, termed N6-methyladenine, protects DNA from restriction endonucleases via the host-specific restriction-modification system. N6-methyladenine was discovered and has been well studied in bacteria. N6-adenine-specific DNA methyltransferase (N6AMT) is the main enzyme catalyzing the methylation of the adenine base and knowledge of this enzyme was mainly derived from work in prokaryotic models. However, large-scale gene discovery at the genome level in many model organisms indicated that the N6AMT gene also exists in eukaryotes, such as humans, mice, fruit flies and plants. Here, we cloned a N6AMT gene from Nilaparvata lugens (Nlu-N6AMT) and amplified its fulllength transcript. Then, we carried out a systematic investigation of N6AMT in 33 publically available insect genomes, indicating that all studied insects had N6AMT. Genomic structure analysis showed that insect N6AMT has short introns compared with the mammalian homologs. Domain and phylogenetic analysis indicated that insect N6AMT had a conserved N6-adenineMlase domain that is specific to catalyze the adenine methylation. Nlu-N6AMT was highly expressed in the adult female. We knocked down Nlu-N6AMT by feeding dsRNA from the second instar nymph to adult female, inducing retard development of adult female. In all, we provide the first genome-wide analysis of N6AMT in insects and presented the experimental evidence that N6AMT might have important functions in reproductive development and ovary maturation.
Abstract Methylation of the N6 position of adenine, termed N6-methyladenine, protects DNA from restriction endonucleases via the host-specific restriction-modification system. N6-methyladenine was discovered and has been well studied in bacteria. N6-adenine-specific DNA methyltransferase (N6AMT) is the main enzyme catalyzing the methylation of the adenine base and knowledge of this enzyme was mainly derived from work in prokaryotic models. However, large-scale gene discovery at the genome level in many model organisms indicated that the N6AMT gene also exists in eukaryotes, such as humans, mice, fruit flies and plants. Here, we cloned a N6AMT gene from Nilaparvata lugens (Nlu-N6AMT) and amplified its fulllength transcript. Then, we carried out a systematic investigation of N6AMT in 33 publically available insect genomes, indicating that all studied insects had N6AMT. Genomic structure analysis showed that insect N6AMT has short introns compared with the mammalian homologs. Domain and phylogenetic analysis indicated that insect N6AMT had a conserved N6-adenineMlase domain that is specific to catalyze the adenine methylation. Nlu-N6AMT was highly expressed in the adult female. We knocked down Nlu-N6AMT by feeding dsRNA from the second instar nymph to adult female, inducing retard development of adult female. In all, we provide the first genome-wide analysis of N6AMT in insects and presented the experimental evidence that N6AMT might have important functions in reproductive development and ovary maturation.
|
|
Received: 18 May 2015
Accepted:
|
| Fund: This work was supported by the National Basic Research Program of China (2012CB114102). |
|
Corresponding Authors:
LI Fei, Tel: +86-571-88982679,E-mail: lifei18@zju.edu.cn
|
| About author: ZHANG Jiao, Tel: +86-25-84399920, E-mail: 1187562476@qq.com; |
Cite this article:
ZHANG Jiao, XING Yan-ru, HOU Bo-feng, YUAN Zhu-ting, LI Yao, JIE Wen-cai, SUN Yang, LI Fei.
2016.
Amplification and function analysis of N6-adenine-specific DNA methyltransferase gene in Nilaparvata lugens. Journal of Integrative Agriculture, 15(3): 591-599.
|
| Adams R, McKay E, Craig L, Burdon R. 1979. Methylationof mosquito DNA. Biochimica et Biophysica Acta (BBA-Nucleic Acids and Protein Synthesis), 563, 72-81Boratyn G M, Camacho C, Cooper P S, Coulouris G, Fong A,Ma N, Madden T L, Matten W T, McGinnis S D, MerezhukY. 2013. BLAST: A more efficient report with usabilityimprovements. Nucleic Acids Research, 41, W29-W33.Chen J, Zhang D, Yao Q, Zhang J, Dong X, Tian H, ZhangW. 2010. Feeding-based RNA interference of a trehalosephosphate synthase gene in the brown planthopper,Nilaparvata lugens. Insect Molecular Biology, 19, 777-786Dohno C, Shibata T, Nakatani K. 2010. Discrimination of N6-methyl adenine in a specific DNA sequence. ChemicalCommunications (Cambridge, United Kingdom), 46,5530-5532Dunn D B, Smith J D. 1955. Occurrence of a new base in thedeoxyribonucleic acid of a strain of Bacterium coli. Nature,175, 336-337Fu Q, Zhang Z, Hu C, Lai F, Sun Z. 2001. A chemically defineddiet enables continuous rearing of the brown planthopper,Nilaparvata lugens (Stål) (Homoptera: Delphacidae).Applied Entomology and Zoology, 36, 111-116 Gasteiger E, Hoogland C, Gattiker A, Wilkins M R, Appel RD, Bairoch A. 2005. Protein identification and analysistools on the ExPASy server. In: The Proteomics ProtocolsHandbook. Springer, New York, USA. pp. 571-607Liu Y, Nie D, Huang Y, Lu G. 2009. RNAi-mediated knock-downof gene mN6A1 reduces cell proliferation and decreasesprotein translation. Molecular Biology Reports, 36, 767-774Nicholas K, Nicholas Jr H, Deerfield D. 1997. GeneDoc:Analysis and visualization of genetic variation. EmbnewNews, 4, 370.Niu Y, Zhao X, Wu Y S, Li M M, Wang X J, Yang Y G. 2013.N6-methyl-adenosine (m6A) in RNA: An old modificationwith a novel epigenetic function. Molecular Biology andEvolution, 11, 8-17Punta M, Coggill P C, Eberhardt R Y, Mistry J, Tate J, BoursnellC, Pang N, Forslund K, Ceric G, Clements J. 2012. ThePfam protein families database. Nucleic Acids Research,40, D290-D301.Ratel D, Ravanat J L, Berger F, Wion D. 2006. N6-methyladenine:The other methylated base of DNA. Bioessays, 28, 309-315Ren L, Zhu B, Zhang Y, Wang H, Li C, Su Y, Ba C. 2004. Theresearch of applying Primer premier 5.0 to design PCRprimer. Journal of Jinzhou Medical College, 25, 43-46.(in Chinese)Schmittgen T D, Livak K J. 2008. Analyzing real-time PCRdata by the comparative CT method. Nature Protocols, 3,1101-1108Shorning B Y, Vanyushin B F. 2001. Putative DNA-(amino)methyltransferases in eucaryotes. Biochemistry (Moscow),66, 753-762Tamura K, Peterson D, Peterson N, Stecher G, Nei M, KumarS. 2011. MEGA5: Molecular evolutionary genetics analysisusing maximum likelihood, evolutionary distance, maximumparsimony methods. Molecular Biology and Evolution, 28,2731-2739Thompson J D, Higgins D G, Gibson T J. 1994. CLUSTAL W:Improving the sensitivity of progressive multiple sequencealignment through sequence weighting, position-specificgap penalties and weight matrix choice. Nucleic AcidsResearch, 22, 4673-4680Vanyushin B F, Ashapkin V V. 2011. DNA methylation in higherplants: past, present and future. Biochimica et BiophysicaActa, 1809, 360-368Wang X, Lu Z, Gomez A, Hon G C, Yue Y, Han D, Fu Y,Parisien M, Dai Q, Jia G, Ren B, Pan T, He C. 2014. N6-methyladenosine-dependent regulation of messenger RNAstability. Nature, 505, 117-120Wion D, Casadesus J. 2006. N6-methyl-adenine: An epigeneticsignal for DNA-protein interactions. Nature ReviewsMicrobiology, 4, 183-192Xue J, Bao Y Y, Li B L, Cheng Y B, Peng Z Y, Liu H, Xu H J, ZhuZ R, Lou Y G, Cheng J A, Zhang C X. 2010. Transcriptomeanalysis of the brown planthopper Nilaparvata lugens. PLoSOne, 5, e14233.Xue J, Zhou X, Zhang C X, Yu L L, Fan H W, Wang Z, Xu HJ, Xi Y, Zhu Z R, Zhou W W, Pan P L, Li B L, ColbourneJ K, Noda H, Suetsugu Y, Kobayashi T, Zheng Y, Liu S,Zhang R, Liu Y, et al. 2014. Genomes of the rice pestbrown planthopper and its endosymbionts reveal complexcomplementary contributions for host adaptation. GenomeBiology, 15, 521. |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
