Transgenic overexpression of UIP1, an interactor of the 3′ untranslated region of the Rubisco small subunit mRNA, increases rice tolerance to drought
- 430 Downloads
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 RNA-binding protein (RBP) possessing a CBS (cystathionine-β-synthase) domain through yeast three-hybrid screening. This RBP, 3′-UTR-interacting protein 1 (UIP1), interacts with 3′ untranslated region of the Rubisco small subunit mRNA (3′ 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. Co-transformation of yeast with these constructs together with three 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 3′ 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 (F v /F m) 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 3′ untranslated region of RbcS1 mRNA and increases tolerance of transgenic overexpressors to drought stress.
KeywordsUIP1 (3′-UTR-interacting protein 1) 3′RbcS Yeast three-hybrid Transgenic rice Drought tolerance SMD
This study was supported by the Rural Development Administration under “Cooperative Research Program for Agriculture Science and Technology Development” (Project No. PJ906910), the Next generation BioGreen 21 Program (Project No. PJ007971 to J.-K.K. and PJ009022 to J.S.J), the Ministry of Education, Science and Technology under “Mid-career Researcher Program” (Project No. 20100026168).
- Castiglioni P, Warner D, Bensen RJ, Anstrom DC, Harrison J, Stoecker M, Abad M, Kumar G, Salvador S, D’Ordine R, Navarro S, Back S, Fernandes M, Targolli J, Dasgupta S, Bonin C, Luethy MH, Heard JE (2008) Bacterial RNA chaperones confer abiotic stress tolerance in plants and improved grain yield in maize under water-limited conditions. Plant Physiol 147:446–455PubMedCrossRefGoogle Scholar
- Hook B, Bernstein D, Zhang B, Wickens M (2005) RNA–protein interactions in the yeast three-hybrid system: affinity, sensitivity, and enhanced library screening. RNA 11:227–233Google Scholar
- Jung H, Kim J-K, Ha S-H (2011) Use of animal viral internal ribosome entry site sequence makes multiple truncated transcripts without mediating polycistronic expression in rice. J Korean Soc Appl Biol Chem 54:678–684Google Scholar
- Kushwaha HR, Singh AK, Sopory SK, Singla-Pareek SL, Pareek A (2009) Genome wide expression analysis of CBS domain containing proteins in Arabidopsis thaliana (L.) Heynh and Oryza sativa L. reveals their developmental and stress regulation. BMC Genomics 10:200. doi: 10.1186/1471-2164-10-200 PubMedCrossRefGoogle Scholar
- Lorković ZJ (2009) Role of plant RNA-binding proteins in development, stress response and genome organization. Trends Plant Sci 14:1360–1385Google Scholar
- Park S-H, Chung PJ, Juntawong P, Bailey-Serres J, Kim YS, Jung H, Bang SW, Kim Y-K, Choi YD, Kim J-K (2012) Post-transcriptional control of photosynthetic mRNA decay under stress conditions requires 3′ and 5′ untranslated regions and correlates with differential polysome association in rice. Plant Physiol. doi: 10.1104/pp.112.194928
- Redillas MCF, Jeong JS, Strasser RJ, Kim YS, Kim J-K (2011) JIP analysis on rice (Oryza sativa cv. Nipponbare) grown under limited nitrogen conditions. J Korean Soc Appl Biol Chem 54(5):827–832Google Scholar
- Wurth L (2012) Versatility of RNA-binding proteins in cancer. Comp Funct Genom. doi: 10.1155/2012/178525