Molecular characterization of two novel molecular chaperones in bacterial-challenged Apostichopus japonicus.
Molecular characterization of two novel molecular chaperones in bacterial-challenged Apostichopus japonicus.
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细菌攻击刺参中两种新型分子伴侣的分子特征。
DOI：
10.1016/j.gene.2015.06.024
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发表时间：
2015-10
期刊：
影响因子：
3.5
通讯作者：
Jin Chunhua
中科院分区：
文献类型：
--
作者：
Zhang Weiwei;Li Chenghua;Lv Zhimeng;Jin Chunhua
78kda葡萄糖调节蛋白（GRP78）和蛋白二硫异构酶（PDI）的分子伴侣参与了蛋白质在内质网（ER）中的折叠和组装。越来越多的证据表明，这两种分子在免疫应答中起着重要作用。本研究利用RNA-seq和RACE方法克隆并鉴定了日本刺参GRP78和PDI基因，分别命名为AjGRP78和AjPDI。AjGRP78 cDNA全长2355 bp，包括一个2013 bp的开放阅读框（ORF），编码一个670个氨基酸的蛋白，具有3个热休克蛋白70 （HSP70）家族特征。AjGRP78在n端含有一个23个氨基酸的信号肽，在c端含有一个HDEL基序，这支持了该蛋白在内质网中的位置。AjPDI cDNA全长1893 bp，其中5 ' untranslation区（UTR） 153 bp, 3 ' untranslation区228 bp， ORF 1512 bp，编码503个氨基酸的蛋白。一个17个氨基酸的信号肽、两个含有两个CGHC活性位点的硫氧还蛋白结构域和KDEL保留信号在AjPDI的推导氨基酸中完全保守。系统发育分析和多重比对表明，这两个基因与其他无脊椎动物和脊椎动物的对应基因具有显著更高的结构保守性和序列一致性，进一步支持这两个蛋白是分子伴侣蛋白家族的新成员。空间表达分析显示，AjGRP78 mRNA转录本主要在触手中表达，而AjPDI mRNA在肌肉、肠道和呼吸树中表达丰富。脾弧菌攻毒海参后，24 h时体腔细胞中AjGRP78和AjPDI mrna的表达量达到峰值，分别是对照组的1.73倍和1.83倍。同样，在1 μg mL−1LPS暴露的原代培养体腔细胞中，AjGRP78和AjPDI的相对mRNA水平也显著升高。这些结果共同提示AjGRP78和AjPDI是a的ER伴侣蛋白。Japonicus，细菌感染诱导表达。
Molecular chaperones of 78 kDa glucose-regulated protein (GRP78) and protein disulfide isomerase (PDI) are involved in protein folding and assembly in the endoplasmic reticulum (ER). Increasing evidences also suggest that these two molecules play an important role in immune response. In the present study, we cloned and characterized GRP78 and PDI genes from Apostichopus japonicus by RNA-seq and RACE approaches (designated as AjGRP78 and AjPDI, respectively). The AjGRP78 cDNA was of 2355 bp including an open reading frame (ORF) of 2013 bp encoding a protein of 670 amino acids with three heat shock protein 70 (HSP70) family signatures. AjGRP78 contained a 23-amino acid signal peptide at the N-terminus and a HDEL motif at the C-terminus, which supported the location of the protein in the ER. The full length cDNA of AjPDI was of 1893 bp with a 5′ untranslated region (UTR) of 153 bp, a 3′ UTR of 228 bp and an ORF of 1512 bp encoding a protein of 503 amino acids. A 17-amino acid signal peptide, two thioredoxin domains with two active sites of CGHC, and KDEL retention signal were totally conserved in the deduced amino acid of AjPDI. Phylogenic analysis and multiple alignments have shown that both genes shared remarkably higher degree of structural conservation and sequence identities with other counterparts from invertebrates and vertebrates, further supporting that the two proteins were novel members of molecular chaperone family. Spatial expression analysis revealed that AjGRP78 mRNA transcripts were dominantly expressed in the tentacle, while AjPDI mRNA levels were abundant in the muscle, intestine and respiratory trees. For Vibrio splendidus challenged sea cucumber, the peak expression of AjGRP78 and AjPDI mRNAs in coelomocytes were detected at 24 h with 1.73-fold increase and at 6 h with 1.83-fold increase compared with the control group, respectively. Similarly, a significant increase in the relative mRNA levels of AjGRP78 and AjPDI was also identified in 1 μg mL− 1LPS exposed primary cultured coelomocytes. These results collectively suggested that AjGRP78 and AjPDI were ER chaperones ofA. japonicus, of which expression is induced upon bacterial infection.
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DOI：
10.1016/j.cbpb.2014.03.001
发表时间：
2014-05
期刊：
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology
影响因子：
--
作者：
Kexue Ma;Guangwen Chen;Changying Shi;F. Cheng;He Dou;C. Feng;Dezeng Liu
通讯作者：
Kexue Ma;Guangwen Chen;Changying Shi;F. Cheng;He Dou;C. Feng;Dezeng Liu
影响因子：
7.3
作者：
Morito D;Nagata K
通讯作者：
Nagata K
真核生物中的氧化蛋白折叠：机制和后果。
DOI：
10.1083/jcb.200311055
发表时间：
2004-02-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Tu BP;Weissman JS
通讯作者：
Weissman JS
线粒体网络和内质网之间的结构和功能联系。
DOI：
10.1016/j.biocel.2009.04.010
发表时间：
2009-10
影响因子：
4
作者：
Giorgi, Carlotta;De Stefani, Diego;Bononi, Angela;Rizzuto, Rosario;Pinton, Paolo
通讯作者：
Pinton, Paolo
DOI：
--
发表时间：
1994-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
H. Lucero;D. Lebeche;B. Kaminer
通讯作者：
H. Lucero;D. Lebeche;B. Kaminer