Molecular Cloning and Expression Analysis of Extra Sex Combs Gene during Development in Macrobrachium nipponense

Year 2013, Volume: 13 Issue: 2, - , 01.04.2013

Yanping Zhang Sufei Jiang Yiwei Xiong Shengming Sun Hui Qiao Shubo Jin Yongsheng Gong Hongtuo Fu

https://doi.org/10.4194/1303-2712-v13_2_15

Abstract

The extra sex combs (Esc) gene first identified in Drosophia encodes a transcriptional repressor of homeotic genes belonging to the polycomb group (PcG) gene and is primarily expressed in the early embryos. In this study, we have isolated the full-length cDNA of an Esc gene from the testis of oriental river prawn (Macmbrachium nipponense) according to the established expressed sequence tags (ESTs) information using Rapid Amplification of the cDNA Ends (RACE) technique, designated as MnEsc. The full-length cDNA of MnEsc is 1,461bp in size and has an open reading frame (ORF) of 1,068bp, encoding a 355-amino acid protein. Real-time quantitative PCR analyses revealed that the expression level of MnEsc varied significantly in the developing embryo, postembryonic larval and adult tissues. Real-time quantitative PCR showed the MnEsc gene was expressed in testis, liver, ovary, brain, abdominal ganglion, heart, intestine and muscle with the highest level of expression in ovary and brain. In vivo silencing of the gene, the dsRNA of MnEsc caused a significantly decrease in target gene expression level in brain and ovary tissues, but no exterior appearance change of experimental shrimp was observed.

Keywords

Macrobrachium nipponense, extra sex combs, real-time quantitative PCR, RNAi

References

• Altschul, S. F., Gish, W., Miller, W., Myers, E. W. and Lipman, D. J. 1990. Basic local alignment search tool. Journal of Molecular Biology, 215: 403-410. doi: 1016/S0022-2836(05)80360-2.
• Bracken, A. P. and Helin, K. 2009. Polycomb group proteins: navigators of lineage pathways led astray in cancer. Nature Reviews Cancer, 9(11):773-784. doi: 1038/nrc2736.
• Breen, T. R. and Duncan, I. M. 1986. Maternal expression of genes that regulate the bithorax complex of Drosophila melanogaster. Developmental Biology, 118: 442-456. doi: 10.1016/0012-1606(86)90015-1.
• Budd, G. E. and Telford, M. J. 2009. The origin and evolution of arthropods. Nature, 457: 812-817. doi: 1038/nature07890.
• Bureau of Fishery, Ministry of Agriculture, P.R.C. 2009. Fisheries economic statistics. In: China Fishery Yearbook. China Agricultural Press, Beijing, 236pp.
• Chen, Y., Zhu, Q., Chen, H., Zhu, X. L., Cui, Z. and Qiu, G. F. 2012. The morphological and histological observation of embryonic development in the oriental river prawn Macrobrachium nipponense. Journal of shanghai ocean university (China), 21(1): 33-40.
• Chiyoko, T. and Curt, S. 1965. The developmental autonomy of extra sex combs in Drosophila melanogaster. Developmental Biology, 11(1): 50-81. doi:1016/0012-1606(65)90037-0
• Duncan, I. M. 1982. Polycomblike: a gene that appears to be required for the normal expression of the bithorax and Antennapedia complexes of Drosophila melanogaster. Genetics, 102: 49http://www.genetics.org/content/102/1/49.
• Dura, J. M., Brock, H. W. and Santamaria, P. 1985. Polyhomeotic: a gene in Drosophila meIanogaster required for correct expression of segment identity. Molecular and General Genetics, 198: 220-231. doi:1007/BF00382998.
• Faust, C., Lawson, K. A., Schork, N. J., Thiel, B. and Magnuson, T. 1998. The Polycomb-group gene eed is required for normal morphogenetic movements during gastrulation in the mouse embryo. Development, 125:4495–4506. http://dev.biologists.org/content/125/22/4495.
• Girton, J. R. and Jeon, S. H. 1994. Novel embryonic and adult homeotic phenotypes are produced by pieiohomeotic mutations in Drosophila. Developmental Biology, 161:393-407. doi:org/10.1006/dbio.1994.1040.
• Gutjahr, T., Frei, E., Spicer, C., Baumgartner, S., White, R. A. H. and Noll, M. 1995. The Polycomb-group gene, extra sex combs, encodes a nuclear member of the WD-40 repeat family. EMBO Journal, 14: 4296– 430 PMCID: PMC394514
• Ingham, P. 1984. A gene that regulates the bithorax complex differentially in larval and adult cells of Drosophila. Cell, 37: 815-823. doi: 10.1016/00928674(84)90416-1.
• Jones, R. S. and Gelbart, W. M. 1990. Genetic analysis of the Enhancer of zeste locus and its role in gene regulation in Drosophila melanogaster. Genetics, 126:185-199. PMCID: PMC1204123.
• Joyce, N. G., Li, R. H., Kelly, M. and Simon, J. 1997. Evolutionary Conservation and Predicted Structure of the Drosophila extra sex combs Repressor Protein. Molecular and cellular biology, 17(11): 6663–6672. doi:0270-7306/97/$04.0010
• JÜrgens, G. 1985. A group of genes controlling the spatial expression of the bithorax complex in Drosophila. Nature, 316:153-155. doi:10.1038/316153a0.
• Katsuhito, O., Donna, M., Birgit, C., Axel, I. and Vincenzo, P. 2008. ESC, ESCL and their roles in Polycomb Group mechanisms, mechanisms of development, 125: 527–541. doi:10.1016/j.mod.2008.01.002.
• Kohler, C. and Hennig, L. 2010. Regulation of cell identity by plant Polycomb and trithorax group proteins. Current Opinion in Genetics & Devlopment, 20(5):541-547. doi: org/10.1016/j.gde.2010.04.015.
• Komachi, K., Redd, M. J. and Johnson, A. D. 1994. The WD repeats of Tup1 interact with the homeo domain protein alpha 2. Genes and Development, 8(23): 285728doi:10.1101/gad.8.23.2857.
• Lewis, E.B. 19 A gene complex controlling segmentation in Drosophila. Nature, 276: 565-570. PMID: 103000.
• Li, Z. Q., Tsuneyuki, T., Kosuke, S., Zhu, L., Xu, J., Hiroaki, M., Jae, M. L. and Takahiro, K. 2012. Identification and characterization of Polycomb group genes in the silkworm, Bombyx mori. Molecular Biology Reports, 39:5575-5588. doi: 10.1007/s11033011-1362-5
• Livak, K. J. and Schmittgen, T. D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (−Delta Delta C (T)) Method. Methods, 25:402–408. doi:org/10.1006/meth.2001.1262.
• Morey, L. and Helin, K. 2010. Polycomb group proteinmediated repression of transcription. Trends in Biochemical Sciences, 35(6):323-332. doi: 1016/j.tibs.2010.02.009.
• Neer, E. J., Schmidt, C. J., Nambudripad, R. and Smith, T. F. 1994. The ancient regulatory-protein family of WD-repeat proteins. Nature, 371: 297-300. doi: 1038/371297a0.
• Norihisa, S., Atsushi, S., Kouji, Y. and Toru, H. 2004. Participation of Polycomb group gene extra sex combs in hedgehog signaling pathway. Biochemical and Biophysical Research Communications, 323: 5235 doi:10.1016/j.bbrc.2004.08.125
• Oktaba, K., Gutierrez, L., Gagneur, J., Girardot, C., Sengupta, A. K., Furlong, E. E. and Muller, J. 2008. Dynamic regulation by Polycomb group protein complexes controls pattern formation and the cell cycle in Drosophila. Developmental Cell, 15(6): 8778 doi:10.1016/j.devcel.2008.10.005.
• Papaceit, M., Antonio, J. S. and Prevosti, A. 1991. Genetic analysis of extra sex combs in the hybrids between Drosophila subobscura and D. madeirensis. Genetics, 84: 107-114.doi: 10.1007/BF00116550.
• Qiao, H., Fu, H. T., Jin, S. B., Wu, Y., Jiang, S. F., Gong, Y. S. and Xiong, Y. W. 2012. Constructing and random sequencing analysis of normalized cDNA library of testis tissue from oriental river prawn (Macrobrachium nipponense).Comparative Biochemistry and Physiology D-genomics & proteomics, 7(3): 268-2 doi:1016/j.cbd. 200003.
• Sathe, S. S. and Harte, P. J. 1995. The Drosophila extra sex combs protein contains WD motifs essential for its function as a repressor of homeotic genes. Mechanisms of Development, 52:77–87. doi:org/10.1016/0925-4773(95)00392-E.
• Sato, T., Hayes, P. H. and Denell, R. E. 1983. Homeosis in Drosophila: maternal effect of the Enhancer of Polycomb locus and its interaction with Polycomb and related loci. Developmental Genetics, 4:185-198. doi: 1002/dvg.1020040305
• Simon, J., Bornemann, D., Lunde, K. and Schwartz, C. 19 The extra sex combs product contains WD40 repeats and its time of action implies a role distinct from other Polycomb group products. Mechanisms of Development, 53: 197-208. doi:org/10.1016/09254773(95)00434-3.
• Struhl, G. 1981. A gene product required for correct initiation of segmental determination in Drosophila. Nature, 293: 36-41. doi:10.1038/293036a0.
• Struhl, G. and Akam, M. 1985. Altered distributions of Ultrabithorax transcripts in extra sex combs mutant embryos of Drosophila. EMBO Journal, 4: 325932 PMCID: PMC554652.
• Struhl, G. and Brower, D. 1982. Early role of the ESC1 gene product in the determination of segments in Drosophila. Cell, 31: 285-292. doi:10.1016/00928674(82)90428-7.
• Tamura, K., Dudley, J., Nei, M. and Kumar, S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 24: 1596-1599. doi: 10.1093/molbev/ msm0 van der Voorn, L. and Ploegh, H. L. 1992. The WD-40 repeat. Febs Letters, 307: 131-1 doi: 1016/0014-5793(92)80751-2.
• Wang, Q., Fang, D.A., Sun, J.L., Wang, Y., Wang, J. and Liu, L.H. 2012. Characterization of the vasa gene in the Chinese mitten crab Eriocheir sinensis: A germ line molecular marker. Journal of Insect Physiology, 58:960–965. doi: 10.1016/j.jinsphys.2012.04.012
• Whiteway, M., Clark, K.L., Leberer, E., Dignard, D. and Thomas, D.Y. 1994. Genetic identification of residues involved in association of α and β G-protein subunits. Molecular and Cellular Biology, 14:3223-3229. doi: 1128/MCB.14.5.3223.
• Zhang, J. S., Evan, B. and Elena, E. 2012. Epigenetic regulation of skin: focus on the Polycomb complex. Cellular and Molecular Life Sciences, 69:2161-2172. doi: 1007/s00018-012-0920-x.