Leclercq Julie, Adams-Phillips Lori C, Zegzouti Hicham, Jones Brian, Latché Alain, Giovannoni James J, Pech Jean-Claude, Bouzayen Mondher
Unité Mixte de Recherche 990, Institut National de la Recherche Agronomique/Institut National Polytechnique-Ecole Nationale Supérieure Agronomique, Boite Postale 107 Auzeville, 31326 Castanet Tolosan cedex, France.
Plant Physiol. 2002 Nov;130(3):1132-42. doi: 10.1104/pp.009415.
LeCTR1 was initially isolated by both differential display reverse transcriptase-polymerase chain reaction screening for tomato (Lycopersicon esculentum) fruit ethylene-inducible genes and through homology with the Arabidopsis CTR1 cDNA. LeCTR1 shares strong nucleotide sequence homology with Arabidopsis CTR1, a gene acting downstream of the ethylene receptor and showing similarity to the Raf family of serine/threonine protein kinases. The length of the LeCTR1 transcribed region from ATG to stop codon (12,000 bp) is more than twice that of Arabidopsis CTR1 (4,700 bp). Structural analysis reveals perfect conservation of both the number and position of introns and exons in LeCTR1 and Arabidopsis CTR1. The introns in LeCTR1 are much longer, however. To address whether this structural conservation is indicative of functional conservation of the corresponding proteins, we expressed LeCTR1 in the Arabidopsis ctr1-1 (constitutive triple response 1) mutant under the direction of the 35S promoter. Our data clearly show that ectopic expression of LeCTR1 in the Arabidopsis ctr1-1 mutant can restore normal ethylene signaling. The recovery of normal ethylene sensitivity upon heterologous expression of LeCTR1 was also confirmed by restored glucose sensitivity absent in the Arabidopsis ctr1-1 mutant. Expression studies confirm ethylene responsiveness of LeCTR1 in various tissues, including ripening fruit, and may suggest the evolution of alternate regulatory mechanisms in tomato versus Arabidopsis.
LeCTR1最初是通过差异显示逆转录酶-聚合酶链反应筛选番茄(Lycopersicon esculentum)果实乙烯诱导基因以及与拟南芥CTR1 cDNA的同源性而分离得到的。LeCTR1与拟南芥CTR1具有很强的核苷酸序列同源性,拟南芥CTR1是一个在乙烯受体下游起作用的基因,与丝氨酸/苏氨酸蛋白激酶的Raf家族具有相似性。LeCTR1从ATG到终止密码子的转录区域长度(12,000 bp)是拟南芥CTR1(4,700 bp)的两倍多。结构分析表明,LeCTR1和拟南芥CTR1的内含子和外显子数量及位置完全保守。然而,LeCTR1中的内含子要长得多。为了探究这种结构保守性是否表明相应蛋白质的功能保守性,我们在35S启动子的指导下,在拟南芥ctr1-1(组成型三重反应1)突变体中表达了LeCTR1。我们的数据清楚地表明,LeCTR1在拟南芥ctr1-1突变体中的异位表达可以恢复正常的乙烯信号传导。拟南芥ctr1-1突变体中不存在的葡萄糖敏感性恢复也证实了LeCTR1异源表达后正常乙烯敏感性的恢复。表达研究证实了LeCTR1在包括成熟果实在内的各种组织中的乙烯反应性,并可能暗示了番茄与拟南芥中替代调控机制的进化。