Interaction Network of the 14-3-3 Protein in the Ancient Protozoan Parasite Giardia duodenalis
Interaction Network of the 14-3-3 Protein in the Ancient Protozoan Parasite Giardia duodenalis
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DOI：
10.1021/pr3000199
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发表时间：
2012-05-01
影响因子：
4.4
通讯作者：
Pozio, Edoardo
中科院分区：
文献类型：
--
作者：
Lalle, Marco;Camerini, Serena;Pozio, Edoardo
14 - 3 - 3s是磷酸丝氨酸/磷酸转铁蛋白结合蛋白，在真核生物中作为多个细胞过程的调节剂发挥关键作用。鞭毛虫寄生虫贾第鞭毛虫是贾第虫病的病原体，是一种有价值的简化真核生物模型。一个单一的14 - 3 - 3同种型（g14 - 3 - 3）在贾第虫中表达，它直接参与寄生虫分化成包囊。为了确定g14 - 3 - 3的总体功能，研究了蛋白质相互作用组。转基因G.将Escherialis菌株工程化以在其自身启动子下表达FLAG标记的g14 - 3 - 3。亲和层析结合串联质谱分析已被用来纯化和鉴定FLAG-g14 - 3 - 3相关蛋白的滋养体和包囊寄生虫。共鉴定了314个推定的g14 - 3 - 3相互作用伴侣，包括参与几种途径的蛋白质。有些互动似乎是特定阶段特有的，而另一些则是不同阶段之间共享的。此外，g14 - 3 - 3与gCDC 7蛋白激酶（gCDC 7）的心血管同源物的相互作用的特点，导致鉴定的多蛋白复合物不仅包含g14 - 3 - 3和gCDC 7，但也是一个新发现的和高度分歧的同源物DBF 4，推定的调控亚基gCDC 7。g14 - 3 - 3相互作用在G.讨论了十二指肠生物学。
14-3-3s are phosphoserine/phosphotreonine binding proteins that play pivotal roles as regulators of multiple cellular processes in eukaryotes. The flagellated protozoan parasite Giardia duodenalis, the causing agent of giardiasis, is a valuable simplified eukaryotic model. A single 14-3-3 isoform (g14-3-3) is expressed in Giardia, and it is directly involved in the differentiation of the parasite into cyst. To define the overall functions of g14-3-3, the protein interactome has been investigated. A transgenic G. duodenalis strain was engineered to express a FLAG-tagged g14-3-3 under its own promoter. Affinity chromatography coupled with tandem mass spectrometry analysis have been used to purify and identify FLAG-g14-3-3-associated proteins from trophozoites and encysting parasites. A total of 314 putative g14-3-3 interaction partners were identified, including proteins involved in several pathways. Some interactions seemed to be peculiar of one specific stage, while others were shared among the different stages. Furthermore, the interaction of g14-3-3 with the giardial homologue of the CDC7 protein kinase (gCDC7) was characterized, leading to the identification of a multiprotein complex containing not only g14-3-3 and gCDC7 but also a newly identified and highly divergent homologue of DBF4, the putative regulatory subunit of gCDC7. The relevance of g14-3-3 interactions in G. duodenalis biology was discussed.