红鳍东方鲀结合CCL25b核心启动子区转录因子的鉴定
Identification of Transcription Factors Binding the Core Promoter Region of CCL25b in Takifugu rubripes
DOI: 10.12677/AMS.2021.84008, PDF,    国家科技经费支持
作者: 尚凤芹:大连海洋大学水产与生命学院,辽宁 大连;大连海洋大学海洋科技与环境学院,辽宁 大连;李 岩, 陆 芸, 王秀利, 刘 洋*:大连海洋大学水产与生命学院,辽宁 大连
关键词: 启动子转录因子DNA pull down质谱鉴定 Promoter Transcription Factor DNA pull down Mass Spectrometry Identification
摘要: 基因转录调控一直是生物信息学研究的一个重要内容,转录因子与启动子区域上的特异性调控元件的结合在基因表达和调控中具有重要意义,是构建基因调控网络的一个核心问题。趋化因子配体25 (chemokine 25, CCL25)是一种属于CC趋化因子家族的小细胞因子,在免疫细胞调节和炎症过程如T细胞归巢和慢性组织炎症中均发挥重要作用。为了探究CCL25在表达过程中受到的转录调控机制,本实验我们初次采用聚合酶链式反应(PCR)扩增出红鳍东方鲀CCL25b的核心启动子区,并利用DNA pull down方法对结合在该序列的蛋白质进行富集,最后结合质谱鉴定数据及GO富集分析初步筛选出6个转录因子(C1QBPPURAARHGAP35NME2RAP2CDRG1),这些初步结果将有助于推进对与CCL25表达调节机制有关的潜在生物学过程的理解。
Abstract: Gene transcriptional regulation has been an important element of bioinformatics research. The binding of transcription factors to specific regulatory elements on promoter regions is important in gene expression and regulation, and is a central issue in the construction of gene regulatory networks. Chemokine ligand 25 (CCL25) is a small cytokine belonging to the CC chemokine family that plays an important role in immune cell regulation and inflammatory processes such as T cell homing and chronic tissue inflammation. In order to investigate the transcriptional regulation mechanism of CCL25 in its expression, Polymerase chain reaction (PCR) was used to amplify the core promoter region of CCL25b in Takifugu rubripes, and then DNA pull down was used to enrich the proteins bound to this sequence. Finally, we combined the mass spectrometry identification data with GO enrichment analysis to initially identify six transcription factors (C1QBP, PURA, ARHGAP35, NME2, RAP2C and DRG1), and these results will help advance the understanding of the underlying biological processes associated with the regulatory mechanism of CCL25 expression.
文章引用:尚凤芹, 李岩, 陆芸, 王秀利, 刘洋. 红鳍东方鲀结合CCL25b核心启动子区转录因子的鉴定[J]. 海洋科学前沿, 2021, 8(4): 69-81. https://doi.org/10.12677/AMS.2021.84008

参考文献

[1] Vicari, A.P., Figueroa, D.J., Hedrick, J.A., Foster, J.S., Singh, K.P., Menon, S., Copeland, N.G., Gilbert, D.J., Jenkins, N.A. and Ba Con, K.B. (1997) TECK: A Novel CC Chemokine Specifically Expressed by Thymic Dendritic Cells and Potentially Involved in T Cell Development. Immunity, 7, 291-301. [Google Scholar] [CrossRef
[2] Ringe, J., Hemmati-Sadeghi, S., Fröhlich, K., Engels, A. and Sittinger, M. (2019) CCL25-Supplemented Hyaluronic Acid Attenuates Cartilage Degeneration in a Guinea Pig Model of Knee Osteoarthritis. Journal of Orthopaedic Research, 37, 1723-1729. [Google Scholar] [CrossRef] [PubMed]
[3] Xu, B.P., Deng, C., Wu, X., Ji, T., Zhao, L., Han, Y.H., Yang, W.W., Qi, Y.T., Wang, Z., Yang, Z., et al. (2020) CCR9 and CCL25: A Review of Their Roles in Tumor Promotion. Journal of Cellular Physiology, 235, 9121-9132. [Google Scholar] [CrossRef] [PubMed]
[4] Cramer, P. (2019) Eukaryotic Transcription Turns 50. Cell, 179, 808-812. [Google Scholar] [CrossRef] [PubMed]
[5] Nagore, L.I., Nadeau, R.J., Guo, Q., Jadhav, Y.L.A., Jarrett, H.W. and Haskins, W.E. (2013) Purification and Characterization of Transcription Factors. Mass Spectrometry Reviews, 32, 386-398. [Google Scholar] [CrossRef] [PubMed]
[6] Ishihama, A. (2000) Functional Modulation of Escherichia Coli RNA Polymerase. Annual Review of Microbiology, 54, 499-518. [Google Scholar] [CrossRef] [PubMed]
[7] Deng, W.G., Ying, Z., Montero, A. and Wu, K.K. (2003) Quantitative Analysis of Binding of Transcription Factor Complex to Biotinylated DNA Probe by a Streptavidin-Agarose Pulldown Assay. Analytical Biochemistry, 323, 12-18. [Google Scholar] [CrossRef] [PubMed]
[8] Tran, D.H., Shishido, Y., Chung, S.P., Trinh, H., Yorita, K., Sakai, T. and Fukui, K. (2015) Identification of DNA-Binding Proteins That Interact with the 5’-Flanking Region of the Human D-Amino Acid Oxidase Gene by Pull-Down Assay Coupled with Two-Dimensional Gel Electrophoresis and Mass Spectrometry. Journal of Pharmaceutical Biomedical Analysis, 116, 94-100. [Google Scholar] [CrossRef] [PubMed]
[9] Schwanhusser, B., Busse, D., Li, N., Dittmar, G., Schuchhardt, J., Wolf, J., Chen, W. and Selbach, M. (2013) Global Quantification of Mammalian Gene Expression Control. Nature, 495, 126-127. [Google Scholar] [CrossRef] [PubMed]
[10] Yu, G., Wang, L.G., Han, Y. and He, Q.Y. (2012) clusterPro-filer: An R Package for Comparing Biological Themes among Gene clusters. Omics—A Journal of Integrative Biology, 16, 284-287. [Google Scholar] [CrossRef] [PubMed]
[11] Wu, K.K. (2006) Analysis of Protein-DNA Binding by Streptavidin-Agarose Pulldown. Methods in Molecular Biology, 338, 281-290. [Google Scholar] [CrossRef] [PubMed]
[12] Bergemann, A.D., Ma, Z.W. and Johnson, E.M. (1992) Sequence of cDNA Comprising the Human Pur Gene and Sequence-Specific Single-Stranded-DNA-Binding Properties of the En-coded Protein. Molecular Cellular Biology, 12, 5673-5682. [Google Scholar] [CrossRef
[13] Hong, L. and Johnson, E.M. (2002) Distinct Proteins Encoded by Alternative Transcripts of the PURG Gene, Located Contrapodal to WRN on Chromosome 8, Determined by Differential Termination/Polyadenylation. Nucleic Acids Research, 30, 2417-2426. [Google Scholar] [CrossRef] [PubMed]
[14] Johnson, E.M. (2003) The Pur Protein Family: Clues to Function from Recent Studies on Cancer and AIDS. Anticancer Research, 23, 2093-2100.
[15] Almut, G., Stéphane, R., Dirk, K., Johannes, S., Dierk, N. and James, H.N. (2010) Of Bits and Bugs—On the Use of Bioinformatics and a Bacte-rial Crystal Structure to Solve a Eukaryotic Repeat-Protein Structure. PLoS ONE, 5, e13402. [Google Scholar] [CrossRef] [PubMed]
[16] Khalili, K., et al. (2003) Pur Is Essential for Postnatal Brain De-velopment and Developmentally Coupled Cellular Proliferation as Revealed by Genetic Inactivation in the Mouse. Molec-ular Cellular Biology, 23, 6857-6875. [Google Scholar] [CrossRef
[17] Hariharan, S., Kelm, R.J. and Strauch, A.R. (2014) The Purα/Purβ Single-Strand DNA-Binding Proteins Attenuate Smooth-Muscle Actin Gene Transactivation in Myofibro-blasts. Journal of Cellular Physiology, 229, 1256-1271. [Google Scholar] [CrossRef] [PubMed]
[18] Kelm, R.J., Elder, P.K., Strauch, A.R. and Getz, M.J. (1997) Sequence of cDNAs Encoding Components of Vascular Actin Single-Stranded DNA-Binding Factor 2 Establish Identity to Puralpha and Purbeta. The Journal of Biological Chemistry, 272, 26727-26733. [Google Scholar] [CrossRef] [PubMed]
[19] Barr, S.M. and Johnson, E.M. (2001) Ras-Induced Colony For-mation and Anchorage-Independent Growth Inhibited by Elevated Expression of Puralpha in NIH3T3 Cells. Journal of Cellular Biochemistry, 81, 621-638. [Google Scholar] [CrossRef] [PubMed]
[20] Liu, H., Barr, S.M., Chu, C., Kohtz, D.S., Kinoshita, Y. and Johnson, E.M. (2005) Functional Interaction of Purα with the Cdk2 Moiety of Cyclin A/Cdk2. Biochemical Biophysical Research Communications, 328, 851-857. [Google Scholar] [CrossRef] [PubMed]
[21] Lupien, S.J., Mcewen, B.S., Gunnar, M.R. and Heim, C. (2009) Effects of Stress throughout the Lifespan on the Brain, Behaviour and Cognition. Nature Reviews Neuroscience, 10, 434-445. [Google Scholar] [CrossRef] [PubMed]
[22] Rhen, T. and Cidlowski, J.A. (2005) Antiinflammatory Action of Glucocorticoids—New Mechanisms for Old Drugs. The New England Journal of Medicine, 353, 1711-1723. [Google Scholar] [CrossRef
[23] Pazirandeh1, A., Xue, Y., Prestegaard, T., Jondal, M. and Okret, S. (2002) Effects of Altered Glucocorticoid Sensitivity in the T Cell Lineage on Thymocyte and T Cell Homeostasis. The FASEB Journal, 16, 727-729. [Google Scholar] [CrossRef] [PubMed]
[24] Postel, E., Berberich, S., Flint, S. and Ferrone, C. (1993) Human c-myc Transcription Factor PuF Identified as nm23-H2 Nucleoside Diphosphate Kinase, a Candidate Suppressor of Tumor Me-tastasis. Science, 261, 478-480. [Google Scholar] [CrossRef] [PubMed]
[25] Dexheimer, T.S., Carey, S.S., Zuohe, S., Gokhale, V.M., Hu, X., Murata, L.B., Maes, E.M., Weichsel, A., Sun, D. and Meuillet, E.J. (2009) NM23-H2 May Play an Indirect Role in Transcriptional Activation of c-myc Gene Expression But Does Not Cleave the Nuclease Hypersensitive Element III1. Molecular Cancer Therapeutics, 8, 1363-1377. [Google Scholar] [CrossRef
[26] Rahl, P.B. and Young, R.A. (2014) MYC and Transcrip-tion Elongation. Cold Spring Harbor Perspectives in Medicine, 4, a020990. [Google Scholar] [CrossRef] [PubMed]
[27] Dominguez-Sola, D., Ying, C.Y., Grandori, C., Ruggiero, L., Chen, B., Li, M., Galloway, D.A., Gu, W., Gautier, J. and Dalla-Favera, R. (2007) Non-Transcriptional Control of DNA Replication by c-Myc. Nature, 448, 445-451. [Google Scholar] [CrossRef] [PubMed]
[28] Guo, Z., Yuan, J., Tang, W., Chen, X., Gu, X., Luo, K., Wang, Y. and Yu, W.L. (2007) Cloning and Characterization of the Human Gene RAP2C, a Novel Member of Ras Family, Which Ac-tivates Transcriptional Activities of SRE. Molecular Biology Reports, 34, 137-144. [Google Scholar] [CrossRef] [PubMed]
[29] Chandrani, C., David, H., Ryuji, K. and Maity, S.N. (2004) Hu-man p32, Interacts with B Subunit of the CCAAT-Binding Factor, CBF/NF-Y, and Inhibits CBF-Mediated Transcription Activation in Vitro. Nucleic Acids Research, 32, 3632-3641. [Google Scholar] [CrossRef] [PubMed]
[30] Huang, L.J., Chi, J., Berry, F., Footz, T.K., Sharp, M.W. and Walter, M.A. (2008) Human p32 Is a Novel FOXC1-Interacting Protein That Regulates FOXC1 Transcriptional Activity in Ocular Cells. Investigative Ophthalmology Visual Science, 49, 5243-5249. [Google Scholar] [CrossRef] [PubMed]
[31] Li, B. and Trueb, B. (2000) DRG Represents a Family of Two Closely Related GTP-Binding Proteins. Biochim Biophys Acta, 1491, 196-204. [Google Scholar] [CrossRef
[32] Leipe, D.D., Wolf, Y.I., Koonin, E.V. and Aravind, L. (2002) Classification and Evolution of P-Loop GTPases and Related ATPases. Journal of Molecular Biology, 317, 41-72. [Google Scholar] [CrossRef] [PubMed]
[33] Markolovic, S., Zhuang, Q., Wilkins, S.E., Eaton, C.D., Ab-boud, M.I., Katz, M.J., McNeil, H.E., Lesniak, R.K., Hall, C., Struwe, W.B., et al. (2018) The Jumonji-C Oxygenase JMJD7 Catalyzes (3S)-lysyl Hydroxylation of TRAFAC GTPases. Nature Chemical Biology, 14, 688-695. [Google Scholar] [CrossRef] [PubMed]
[34] Perez-Arellano, I., Spinola-Amilibia, M. and Bravo, J. (2013) Human Drg1 Is a Potassium-Dependent GTPase Enhanced by Lerepo4. FEBS Journal, 280, 3647-3657. [Google Scholar] [CrossRef] [PubMed]