Characterization of two novel subtilases from common bean (Phaseolus vulgaris L.) and their responses to drought
Characterization of two novel subtilases from common bean (Phaseolus vulgaris L.) and their responses to drought
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DOI：
10.1016/j.plaphy.2012.10.022
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发表时间：
2013-01-01
影响因子：
6.5
通讯作者：
Kidric, Marjetka
中科院分区：
文献类型：
--
作者：
Budic, Maruska;Sabotic, Jerica;Kidric, Marjetka
蛋白酶降解蛋白质是植物对干旱等非生物胁迫响应的基础。植物中存在大量编码蛋白酶或推定蛋白酶的基因。只有少数参与应对干旱的生物体得到了鉴定，对它们的调节也知之甚少。从菜豆叶片中鉴定出两种新的枯草杆菌酶。栽培品种Zorin、PvSLP 1和PvSLP 2。根据花生（Arachis hypogaea L.）干旱诱导丝氨酸蛋白酶基因序列设计引物，在基因水平上对PvSLP 1进行了鉴定。在寻常疟原虫中，PvSLP 1转录物的表达在脱水时没有变化。分离PvSLP 2，并在蛋白质水平上进行表征，以及完整的基因和cDNA序列。PvSLP 1和PvSLP 2推导的氨基酸序列都是SB家族S8家族植物枯草杆菌酶的特征。PvSLP 2与PvSLP 1具有33%的序列同一性。PvSLP 2转录的表达没有改变撤回的水，但其蛋白水解活性在叶中增加，这取决于叶的年龄和位置。此外，在水分充足的植物衰老叶片中的活性水平高于成熟或年轻的叶片。这些结果，连同PvSLP 2切割精氨酸残基后的肽键的事实，指出PvSLP 2枯草杆菌酶活性在翻译和/或翻译后水平的调节，并建议在响应干旱和衰老的特定作用。(c)2012年Elsevier Masson SAS。All rights reserved.
Protein breakdown by proteases is basic to the plant response to abiotic stresses such as drought. A large number of genes encoding proteases or putative proteases exist in plants. Only a few of those involved in the response to drought have been characterized, and their regulation is poorly understood. We have identified two new subtilases from leaves of Phaseolus vulgaris L. cultivar Zorin, PvSLP1 and PvSLP2. PvSLP1 was identified at the gene level, using primers based on the gene sequence of the putative drought induced serine protease from Arachis hypogaea L. In P. vulgaris, expression of the PvSLP1 transcript did not change on water withdrawal. PvSLP2 was isolated and characterized at the protein level, together with complete gene and cDNA sequences. The deduced amino acid sequences of both PvSLP1 and PvSLP2 are characteristic of plant subtilases of the S8 family of clan SB. PvSLP2 shows 33% sequence identity to PvSLP1. Expression of the PvSLP2 transcript did not change on withdrawal of water, but its proteolytic activity in leaves increased, depending on the age and position of the leaf. In addition, the level of activity in senescent leaves of well watered plants was higher than in mature or young leaves. These results, together with the fact that PvSLP2 cleaves peptide bonds following an Arg residue, point to regulation of PvSLP2 subtilase activity at translational and/or post-translational levels and suggest a specific role in the response to drought and senescence. (c) 2012 Elsevier Masson SAS. All rights reserved.