A novel protein from Brassica napus has a putative KID domain and responds to low temperature
A novel protein from Brassica napus has a putative KID domain and responds to low temperature
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DOI：
10.1046/j.1365-313x.2003.01694.x
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发表时间：
2003-03-01
期刊：
影响因子：
7.2
通讯作者：
Hannoufa, A
中科院分区：
文献类型：
--
作者：
Gao, MJ;Schäfer, UA;Hannoufa, A
为了鉴定与甘蓝型油菜中组蛋白脱乙酰酶 (HDAC) 相互作用的因子，使用拟南芥 HDA19 作为诱饵筛选了酵母双杂交文库。一种新的蛋白质 bnKCP1，含有假定的激酶诱导结构域 (KID)，被发现与 HDA19 相互作用。 Southern印迹分析表明bnKCP1基因属于一个至少有三个成员的小基因家族。 Northern印迹分析显示bnKCP1在茎、花、根和未成熟的长角果中强烈表达，但在幼苗的叶片中不表达。欧洲油菜幼苗暴露于冷胁迫后 4 小时内，叶片中 bnKCP1 转录物的积累被显着诱导，而用 Inomycin（一种 Ca2+ 离子载体）处理叶片，引起 bnKCP1 表达的快速（30 分钟）但短暂的诱导。与叶片中观察到的相反，茎中 bnKCP1 的表达在冷处理后受到抑制。体外和体内蛋白质结合测定表明，bnKCP1 通过 KID 结构域与 HDA19 相互作用，而 S-188 对于 bnKCP1-HDA19 相互作用至关重要。 BnKCP1 还通过其 N 末端区域对酵母中的 lac Z 报告基因进行适度的反式激活。这些测定表明 bnKCP1 可能作为转录因子发挥作用，通过与 HDA19 相互作用来调节基因表达。
To identify factors that interact with histone deacetylase (HDAC) in Brassica napus , a yeast two-hybrid library was screened using the Arabidopsis HDA19 as bait. A novel protein, bnKCP1, containing a putative kinase-inducible domain (KID) was found to interact with HDA19. Southern blot analysis indicated that the bnKCP1 gene belongs to a small gene family of at least three members. Northern blot analysis showed bnKCP1 to be strongly expressed in stems, flowers, roots, and immature siliques, but not in leaf blades of seedlings. The accumulation of bnKCP1 transcript in the leaf blades was induced significantly within 4 h of exposure of B. napus seedlings to cold stress, whereas treatment of leaf blades with inomycin, an ionophore of Ca2+, caused a rapid (30 min) but transient induction of bnKCP1 expression. In contrast to that observed in leaf blades, expression of bnKCP1 in the stems was repressed upon cold treatment. In vitro and in vivo protein-binding assays showed that bnKCP1 interacts with HDA19 via the KID domain, and that S-188 is critical for bnKCP1-HDA19 interaction. BnKCP1 also exerted modest transactivation of the lac Z reporter gene in yeast through its N-terminal region. These assays suggest that bnKCP1 may function as a transcription factor, which regulates gene expression through interaction with HDA19.