Transgenic overexpression of UIP1, an interactor of the 30 untranslated region of the Rubisco small subunit mRNA, increases rice tolerance to drought

다국어 초록 (Multilingual Abstract)

Gene regulation at the post-transcriptional level is a well-organized process to adjust plants in response to environmental changes. Here, we identified a novel RNAbinding protein (RBP) possessing a CBS (cystathionine-bsynthase) domain through yeast three-hybrid screening. This RBP, 30-UTR-interacting protein 1 (UIP1), interacts with 30 untranslated region of the Rubisco small subunit mRNA (30 RbcS)—the major mRNA element that mediates the stress-induced mRNA decay (SMD) under drought and salt stress conditions. Six deletion constructs were made to delineate the binding domain of the UIP1 protein. Cotransformation of yeast with these constructs together withthree different hybrid RNAs in various combinations showed that deletion of 51 N-terminal amino acids resulted in a loss of sequence-specific binding affinity. Further deletion at the region between 52 and 212 amino acids revealed that the CBS domain of UIP1 is necessary for binding to 30 RbcS. Transgenic overexpression of UIP1 in rice resulted in an increase in tolerance to drought stress at the vegetative stage of growth. Under drought, high salt and low temperature conditions, the maximum photochemical efficiency of photosystem II (Fv/Fm) of UIP1 plants was higher than those of the nontransgenic plants. Interestingly, the effect of UIP1 overexpression on tolerance to stress was much more pronounced under drought than under high salt and low temperature conditions. Taken together, our results demonstrate that UIP1 interacts with 30 untranslated region of RbcS1 mRNA and increases tolerance of transgenic overexpressors to drought stress.

참고문헌 (Reference)

1 Wurth L, "Versatility of RNA-binding proteins in cancer" 2012

2 Harin Jung, "Use of Animal Viral Internal Ribosome Entry Site Sequence Makes Multiple Truncated Transcripts without Mediating Polycistronic Expression in Rice" 한국응용생명화학회 54 (54): 678-684, 2011

3 Lee BH., "STABILIZED1, a stress upregulated nuclear protein, is required for pre-mRNA splicing, mRNA turnover, and stress tolerance in Arabidopsis" 18 : 1736-1749, 2006

4 Hedbacker K., "SNF1/AMPK pathways in yeast" 13 : 2408-2420, 2009

5 Jeong JS., "Root specific expression of OsNAC10 improves drought tolerance and grain yield in rice under field drought conditions" 153 : 185-197, 2010

6 Lorkovic´ ZJ, "Role of plant RNA-binding proteins in development, stress response and genome organization" 14 : 1360-1385, 2009

7 Morris RT., "RiceRBP : a database of experimentally identified RNA-binding proteins in Oryza sativa L" 180 : 204-211, 2011

8 Hook B., "RNA–protein interactions in the yeast three-hybrid system : affinity, sensitivity, and enhanced library screening" 11 : 227-233, 2005

9 Xu J., "Processing bodies and plant development" 14 : 88-93, 2011

10 Park S-H., "Post-transcriptional control of photosynthetic mRNA decay under stress conditions requires 30 and 50 untranslated regions and correlates with differential polysome association in rice" 2012

11 Guthrie C., "Nucleus and gene expression : coordinated nuclear events regulate mRNA synthesis, processing, export and turnover" 17 : 239-241, 2005

12 Meyer S., "Nucleotide recognition by the cytoplasmic domain of the human chloride transporter ClC-5" 14 : 60-67, 2007

13 Dreyfuss G., "Messenger-RNA-binding proteins and the messages they carry" 3 : 195-205, 2002

14 Lejeune F., "Mechanistic links between nonsensemediated mRNA decay and pre-mRNA splicing in mammalian cells" 17 : 309-315, 2005

15 Mark Christian Felipe Reveche Redillas, "JIP Analysis on Rice (Oryza sativa cv Nipponbare) Grown under Limited Nitrogen Conditions" 한국응용생명화학회 54 (54): 827-832, 2011

16 Zhu J., "Interplay between cold-responsive gene regulation, metabolism and RNA processing during plant cold acclimation" 10 : 290-295, 2007

17 Matsumoto K., "Hsp90 is involved in the formation of P-bodies and stress granules" 407 : 720-724, 2011

18 Jang I-C., "High-Level and ubiquitous expression of the rice cytochrome c gene OsCc1 and its promoter activity in transgenic plants provides a useful promoter for transgenesis of monocots" 129 : 1473-1481, 2002

19 Kim JY., "Glycine-rich RNA-binding proteins are functionally conserved in Arabidopsis thaliana and Oryza sativa during cold adaptation process" 61 : 2317-2325, 2010

20 Bailey-Serres J., "Getting the message across : cytoplasmic ribonucleoprotein complexes" 14 : 443-453, 2009

21 Kushwaha HR., "Genome wide expression analysis of CBS domain containing proteins in Arabidopsis thaliana (L.) Heynh and Oryza sativa L. reveals their developmental and stress regulation" 10 : 200-, 2009

22 Lorkovic´ ZJ., "Genome analysis : RNA recognition motif(RRM)and K homology(KH)domain RNA-binding proteins from the flowering plant Arabidopsis thaliana" 30 : 623-635, 2002

23 Este´vez R., "Functional and structural conservation of CBS domains from CLC chloride channels" 557 : 363-378, 2004

24 Moore MJ, "From birth to death : the complex lives of mRNAs" 309 : 1514-1518, 2005

25 Hiei Y., "Efficient transformation of rice(Oryza sativa L.)mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA" 6 : 271-282, 1994

26 Castiglioni P., "Bacterial RNA chaperones confer abiotic stress tolerance in plants and improved grain yield in maize under water-limited conditions" 147 : 446-455, 2008

27 Bernstein DS., "Analyzing mRNA–protein complexes using a yeast three-hybrid system" 26 : 123-141, 2002

28 Hardie DG, "AMP-activated/SNF1 protein kinases : conserved guardians of cellular energy" 8 : 774-785, 2007

29 SenGupta DJ., "A three-hybrid system to detect RNA–protein interactions in vivo" 93 : 8496-8501, 1996

30 Gong Z., "A dead box RNA helicase is essential for mRNA export and important for development and stress responses in Arabidopsis" 17 : 256-267, 2005