Heterologous Expression of a Salinity and Developmentally Regulated Rice Cyclophilin Gene (OsCyp2) in E-coli and S-cerevisiae Confers Tolerance Towards Multiple Abiotic Stresses
Heterologous Expression of a Salinity and Developmentally Regulated Rice Cyclophilin Gene (OsCyp2) in E-coli and S-cerevisiae Confers Tolerance Towards Multiple Abiotic Stresses
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DOI：
10.1007/s12033-009-9153-0
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发表时间：
2009-06-01
影响因子：
2.6
通讯作者：
Pareek, Ashwani
中科院分区：
文献类型：
--
作者：
Kumari, Sumita;Singh, Prabhjeet;Pareek, Ashwani
亲环素2（Cyclophilin 2，OsCyp 2）是水稻亲免素家族的胞质成员。我们已经分离了它的全长cDNA（1，056 bp），其开放阅读框为519 bp，编码172个氨基酸的多肽，估计的pI为8.61。以N-琥珀酰-ala-ala-pro-phe-p-nitroanilidine为肽底物测定了该蛋白的肽基脯氨酰顺反异构酶活性。它的催化效率（K（cat）/K（m））为4.5 × 10（6）/（mol/l）/s，与已知的植物亲环素相当。其活性被亲环素的大环内酯类药物抑制剂环孢菌素A特异性抑制。转录本分析表明，该基因是一个发育差异调控基因，在非胁迫和盐胁迫条件下，在苗期、分蘖期和抽穗期表达量均发生变化。OsCyp 2的表达增强了大肠杆菌在多种非生物胁迫下的生存能力，即盐、高温、渗透胁迫（甘露醇）和氧化胁迫（H2 O2）。OsCyp 2能够补充缺乏天然Cyp 2的酵母突变体，并且还改善了野生型酵母在上述胁迫条件下的生长。基于这些结果，我们建议OsCyp 2可以作为一个“合适的候选人”提高转基因植物增强多重非生物胁迫的耐受性。
Cyclophilin 2 (OsCyp2) is a cytosolic member of immunophilin family from rice. We have isolated its full length cDNA (1,056 bp) with an open reading frame of 519 bp encoding a polypeptide of 172 amino acids and an estimated pI of 8.61. Peptidyl prolyl cis-trans isomerase activity of the protein was determined using N-succinyl-ala-ala-pro-phe-p-nitroanilidine as peptide substrate. It has a catalytic efficiency (K (cat)/K (m)) of 4.5 x 10(6)/(mol/l)/s, which is comparable to known cyclophilins from plants. Its activity is specifically inhibited by cyclosporin A, a macrolide drug inhibitor of cyclophilins. Transcript analysis showed it to be a developmentally and differentially regulated gene; showing changes in abundance at seedling, tillering and heading stage under non-stress and salinity stress conditions. Expression of OsCyp2 enhances the ability of Escherichia coli to survive under diverse abiotic stresses viz. salinity, high temperature, osmotic stress (mannitol) and oxidative stress (H2O2). OsCyp2 was able to complement the yeast mutant lacking native Cyp2 and also improved the growth of wild type yeast under above-mentioned stress conditions. Based on these results, we propose that OsCyp2 may serve as a 'suitable candidate' for raising transgenic plants for enhanced multiple abiotic stress tolerance.