Cloning and Sequence Analysis of Fraxinus velutina smh1 Gene Coding Region

doi:10.11924/j.issn.1000-6850.casb16120077

Abstract

Abstract: To study the function of Fraxinus velutina smh1 telomere binding protein gene in telomere and regulation mechanismin in the process of plant growth and development, we clone and analysis Fraxinus velutina smh1 telomere binding protein gene. By selecting Fraxinus velutina seedling as experiment material and based on the transcriptome Unigene data which gained by Illumina HiSeq 2000 high throughput sequencing technology, this experiment choose RT-PCR method to clone Fraxinus velutina smh1 Gene Coding Region and to make bioinformatics analysis. The results showed that a sequence of Fraxinus velutina smh gene coding region was cloned successfully. Gene coding region length is 879bp, encoding 292 amino acids with a mass of 31.66 kDa, pI of 6.5, atoms to the total number4457, molecular formula C1371H2242N392O439S13. Conserved Domains database analyzed the protein conserved domains and the result shows that this protein sequence have the typical structures of plant Smh telomere binding protein subfamily including the Myb like domain located at the N-terminal and the middle histone H1/5 like domain. Carrying on constructing phylogenetic trees between FvSmh1 and other telomere binding protein which contain telobox shows that both SMH telomere binding protein and TRF1, TRF2 belong to a same branch and have close relationship with each other. From the analysis above, it is inferred that the cloned Fraxinus velutina smh1 gene may play an important role in maintaining the structure and function of the plant telomere.

CLC Number:

S722.5

Zhang Dongjie,Yan Liping,Wu Qichao,Liu Cuilan,Wang Yinhua and Wu Dejun. Cloning and Sequence Analysis of Fraxinus velutina smh1 Gene Coding Region[J]. Chinese Agricultural Science Bulletin, 2017, 33(15): 49-55.

References

[1] Richards E J, Ausubel F M. Isolation of a higher eukaryotic telomere from Arabidopsis thaliana[J]. Cell,1988,53(1):127-136.
[2] 童夏静,周金秋.端粒和端粒酶的发现及其生物学意义[J].生命科学,2009,21(6).
[3] Lange T D. Shelterin: the protein complex that shapes and safeguards human telomeres[J]. Genes & Development,2005,19(18): 2100-2110.
[4] Lange T D. How Telomeres Solve the End-Protection Problem[J]. Science, 2009,326(5955):948-52.
[5] 陈清,汤浩茹,董晓莉,等.植物Myb转录因子的研究进展[J].基因组学与应用生物学,2009,28(2):365-372.
[6] Karamysheva Z N, Surovtseva Y V, Vespa L, et al. A C-terminal Myb extension domain defines a novel family of double-strand telomeric DNA-binding proteins in Arabidopsis[J]. Journal of Biological Chemistry,2004,279(46):47799-47807.
[7] Yu E Y, Kim S E, Kim J H, et al. Sequence-specific DNA recognition by the Myb-like domain of plant telomeric protein RTBP1[J]. Journal of Biological Chemistry,2000,275(31):24208-24214.
[8] Yang S W, Kim D H, Lee J J, et al. Expression of the telomeric repeat binding factor gene NgTRF1 is closely coordinated with the cell division program in tobacco BY-2 suspension culture cells[J]. Journal of Biological Chemistry,2003,278(24):21395-21407.
[9] Moriguchi R, Kanahama K, Kanayama Y. Characterization and expression analysis of the tomato telomere-binding protein LeTBP1[J]. Plant Science,2006,171(1):166-174.
[10] Pe?ka V, Schrumpfová P, Fajkus J. Using the Telobox to Search for Plant Telomere Binding Proteins[J]. Current Protein & Peptide Science,2011,12(2):75-83.
[11] Hwang M G, Chung I K, Kang B G, et al. Sequence specific binding property of Arabidopsis thaliana telomeric DNA binding protein 1 (AtTBP1)[J]. FEBS Letters,2001,503(1):35-40.
[12] Chen C M, Wang C T, Ho C H. A plant gene encoding a Myb-like protein that binds telomeric GGTTTAG repeats in vitro[J]. Journal of Biological Chemistry,2001,276(19):16511-16519.
[13] Marian C O, Bordoli S J, Goltz M, et al. The maize Single myb histone 1 gene,Smh1, belongs to a novel gene family and encodes a protein that binds telomere DNA repeats in vitro[J]. Plant Physiology,2003,133(3):1336-1350.
[14] Byun M Y, Hong J P, Kim W T. Identification and characterization of three telomere repeat-binding factors in rice[J]. Biochemical & Biophysical Research Communications,2008,372(1):85-90.
[15] Schrumpfova P, Kuchar M, Mikova G, et al. Characterization of two Arabidopsis thalianamyb-like proteins showing affinity to telomeric DNA sequence[J]. Genome,2004,47:316-324.
[16] Dvorackova M, Rossignol P, Shaw P J, et al. AtTRB1, a telomeric DNA-binding protein from Arabidopsis, is concentrated in the nucleolus and shows highly dynamic association with chromatin[J]. Plant Journal,2010,61(4):637-649.
[17] Hong J P, Byun M Y, Koo D H, et al. Suppression of RICE TELOMERE BINDING PROTEIN1 Results in Severe and Gradual Developmental Defects Accompanied by Genome Instability in Rice[J]. Plant Cell,?2007,19(6):1770-1781.
[18] Yang S W, Kim S K, Kim W T. Perturbation of NgTRF1 expression induces apoptosis-like cell death in tobacco BY-2 cells and implicates NgTRF1 in the control of telomere length and stability[J]. Plant Cell,2005,16(12): 3370-3385.
[19] Fulcher N, Riha K. Using centromere mediated genome elimination to elucidate the funcional redudancy of candidate telomere binding proteins in Arabidopsis thaliana. Frontiers in Genetics,2016,6:349.
[20] Mozgová I, Schrumpfová P P, Hofr C, et al. Functional characterization of domains in AtTRB1, a putative telomere-binding protein in Arabidopsis thaliana[J]. Phytochemistry,2008,69(9):1814-1819.
[21] Schrumpfová P P, Kuchar M, Palecek J, et al. Mapping of interaction domains of putative telomere-binding proteins AtTRB1 and AtPOT1b from Arabidopsis thaliana[J]. FEBS Letters,2008,582(10):1400-1406.
[22] Schrumpfová P P, Vychodilová I, Dvo?á?ková M, et al. Telomere repeat binding proteins are functional components of Arabidopsis telomeres and interact with telomerase[J]. Plant Journal for Cell & Molecular Biology,2014,77(5):770-781.
[23] Schrumpfová P P, Scho?ová S, Fajkus J. Telomere- and Telomerase- Associated Proteins and Their Functions in the Plant Cell[J]. Frontiers in Plant Science,2016,7(188).
[24] 刘佳,徐秉良,薛应钰,等. 美洲南瓜(Cucurbita pepo)种皮苯丙氨酸解氨酶基因克隆与表达分析[J].中国农业科学,2014,47(6):1216-1226.
[25] Bilaud T, Koering C E, Binet-Brasselet E, et al. The telobox, a Myb- related telomeric DNA binding motif found in proteins from yeast, plants and human[J]. Nucleic Acids Research,1996,24(7):1294-1303.
[26] Chong L, van Steensel B, Broccoli D, et al. A human telomeric protein[J]. Science, 1995,270(5242):1663-1637.
[27] Broccoli D, Smogorzewska A, Chong L, et al. Human telomeres contain two distinct Myb-related proteins, TRF1 andTRF2[J]. Nature Genetics,1997,17(2): 231-235.
[28] Spallarossa P, Altieri P, Aloi C, et al. Doxorubicin induces senescence or apoptosis in rat neonatal cardiomyocytes by regulating the expression levels of the telomere binding factors 1 and 2[J]. Am J Physiol Heart Circ Physiol,2009,297(6):1-6.
[29] Cooley C, Baird K M, Faure V, et al. Trf1 is not required for proliferation or functional telomere maintenance in chicken DT40 cells[J]. Molecular Biology of the Cell,2009,20(10):2563-2571.
[30] Smogorzewska A, Steensel B V, Bianchi A, et al. Control of Human Telomere Length by TRF1 and TRF2[J]. Molecular & Cellular Biology,2000,20(5):1659-1668.
[31] Hwang M G, Cho M H. Arabidopsis thaliana telomeric DNA-binding protein 1 is required for telomere length homeostasis and its Myb-extension domain stabilizes plant telomeric DNA binding[J]. Nucleic Acids Research, 2007,35(4):1333-1342.
[32] Sue S C, Hsiao H H, Chung B C, et al. Solution structure of the Arabidopsis thaliana telomeric repeat-binding protein DNA binding domain: a new fold with an additional C-terminal helix[J]. Journal of Molecular Biology,2006,356(1):72-85.