Abstract
Rice (Oryza sativa) ClpB-C (OsClpB-C) protein is expressed upon heat stress in vegetative tissues and constitutively in seeds. We produced stably transformed Arabidopsis plants carrying β-glucuronidase (Gus) reporter gene downstream to 1-kb OsClpB-C promoter (1kbPro plants). In the 1kbPro plants, expression of Gus transcript and protein followed the expression pattern of OsClpB-C gene in rice plants, i.e., heat induced in vegetative tissues and constitutive in seeds. Next, we produced transgenic Arabidopsis plants containing Gus downstream to 862-bp fragment of OsClpB-C promoter [lacking 138 nucleotides from 3′ end of the 5′untranslated region (5′UTR); ∆UTR plants). In ∆UTR plants, Gus transcript was expressed in heat-inducible manner, but strikingly, Gus protein levels were negligible after heat treatment. However, Gus protein was expressed in ∆UTR seedlings at levels comparable to 1kbPro seedlings when recovery treatment of 22 °C/2 h was given post heat stress (38 °C/15 min). This suggests that 5′UTR of OsClpB-C gene is involved in its post-transcriptional regulation and is an obligate requirement for protein expression during persistent heat stress. Furthermore, the Gus transcript levels were higher in the polysomal RNA fraction in heat-stressed seedlings of 1kbPro plants as compared to ∆UTR plants, indicating that 5′UTR aids in assembly of ribosomes onto the Gus transcript during heat stress. Unlike the case of seedlings, Gus protein was formed constitutively in ∆UTR seeds at levels comparable to 1kbPro seeds. Hence, the function of 5′UTR of OsClpB-C is dispensable for expression in seeds.
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Acknowledgments
RCM, AS, and LDT acknowledge the Council of Scientific and Industrial Research (CSIR), the Government of India, New Delhi, for the Fellowship awards. Richa acknowledges the Department of Science and Technology, the Government of India, for the Inspire Fellowship Award. AG acknowledges the J.C. Bose Fellowship award grant from the Department of Science and Technology (DST), the Government of India, and financial support from the Center for Advanced Research and Innovation on Plant Stress and Developmental Biology, Department of Biotechnology (DBT), Government of India.
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Mishra, R.C., Richa, Singh, A. et al. Characterization of 5′UTR of rice ClpB-C/Hsp100 gene: evidence of its involvement in post-transcriptional regulation. Cell Stress and Chaperones 21, 271–283 (2016). https://doi.org/10.1007/s12192-015-0657-1
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DOI: https://doi.org/10.1007/s12192-015-0657-1