Cloning, Expression and Function Preliminary Research of RPS14 in Arabidopsis thaliana

Abstract

Abstract: RPS14(Ribosomal protein S14)is a ribosomal protein of Arabidopsis thaliana. It is a structural component of 40S ribosomalsubunit and belongs to the S11P ribosomal protein family. It is responsible for RNA post-transcriptional processing. We extracted the total RNAof Arabidopsis thaliana. The RPS14 full-length gene was obtained by RT-PCR. Through inserting the gene fragment into pEASY-T3 vector byT-A cloning, we successfully constructed a recombinant vector pGEX-6P-1-RPS14. The GST-RPS14 protein was expressed in BL21(DE3)strain and molecular weight was approximately 43 kD. We constructed the recombinant vector pET-28a-AK6 in the same way and obtained theapproximately 26 kD HIS-AK6 protein. We demonstrated the interaction between RPS14 and AK6 using pull-down, SDS-PAGE and Westernblot. Therefore, we speculate that they together participate in the nucleic acid metabolism and play a role in the RNA post-transcriptionalprocessing, thereby inhibiting growth of stem cells and making plants show dwarf phenotype.

Key words: Arabidopsis thaliana , RPS14 , AK6 , Pull-down

Fan Yanrong, Liu Ying, Zhang Feiyun. Cloning, Expression and Function Preliminary Research of RPS14 in Arabidopsis thaliana[J]. Biotechnology Bulletin, 2013, 0(1): 73-77.

References

[1] Granneman S, Nandineni MR, Baserga SJ. The Putative NTPaseFap7 mediates cytoplasmic 20S pre-rRNA processing through adirect interaction with RpS14[J]. Mol Cell Biol, 2005, 25（23）：10352-10364.
[2] Whitford PC, Miyashita O, Levy Y, Onuchic JN. Conformationaltransitions of adenylate kinase ：Switching by cracking[J]. J MolBiol, 2007, 366（5）：1661-1671.
[3] Van Rompay AR, Johansson M, Karlsson A. Identifucation of a novelhuman adenylate kinase. cDNA cloning, expression analysis, chromosomelocalization and characterization of the recombinant protein[J]. Eur J Biochem, 1999, 261 ：509-517.
[4] Zancani M, CasoloV, VianelloA, Macri F. Involvement of apyrasein the regulation of the adenylate pool by adenylate kinase in plantmitochondria[J]. Plant Sci, 2001, 161（5）：927-933.
[5] Zhai R, Meng G, Zhao Y, et al. A novel nuclear-localized proteinwith special adenylate kinase properties from Caenorhabditiselegans[J]. FEBS Lett, 2006, 580（16）：3811-3817.
[6] Ren H, Wang L, Bennett M, et al. The crystal structure of humanadenylate kinase 6 ：An adenylate kinase localized to the cell nucleus[J]. Proc Natl Acad Sci, 2005, 102（2）：303-308.
[7] Lange PR, Geserick C, Tischendorf G, Zrenner R. Functions of chloroplasticadenylate kinases in Arabidopsis[J]. Plant Physiology,2007, 146（2）：492-504.
[8] Juhnke H, Charizanis C, Latifi F, et al. The essential protein fap7is involved in the oxidative stress response of Saccharomycescerevisiae[J]. Mol Microbiol, 2000, 35（4）：936-948.
[9] Carrari F, Coll-Garcia D, Schauer N, et al. Deficiency of a plastidialadenylate kinase in Arabidopsis results in elevated photosyntheticamino acid biosynthesis and enhanced growth[J]. PlantPhysiologl, 2005, 137（1）：70-82.
[10] Zhang J, Zhang F, Zheng X. Depletion of hCINAP by RNAinterference causes defects in Cajal body formation, histonetranscription, and cell viability[J]. Cellular and Molecular LifeSciences, 2010, 67（11）：1907-1918.
[11] Meng G, Zhai R, Chen B, et al. Identification and characterization ofhuman adenylate kinase 6 homologues from Drosophila melanogaster[J]. Biochemistry（Moscow）, 2008, 73（1）：38-43.

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