PUBLICATION
Grass carp (Ctenopharyngodon idella) ATF6 (Activating Transcription Factor 6) modulates the transcriptional level of GRP78 and GRP94 in CIK cells
- Authors
- Wang, X., Zhang, T., Mao, H., Mi, Y., Zhong, B., Wei, L., Liu, X., Hu, C.
- ID
- ZDB-PUB-160319-11
- Date
- 2016
- Source
- Fish & shellfish immunology 52: 65-73 (Journal)
- Registered Authors
- Keywords
- ATF6, Endoplasmic Reticulum stress, GRP78 and GRP94, Transcriptional control, Unfolded protein respond
- MeSH Terms
-
- Activating Transcription Factor 6/chemistry
- Activating Transcription Factor 6/genetics*
- Activating Transcription Factor 6/metabolism*
- Amino Acid Sequence
- Animals
- Base Sequence
- Carps/genetics*
- Carps/immunology*
- Carps/metabolism
- Cell Line
- Fish Proteins/chemistry
- Fish Proteins/genetics*
- Fish Proteins/metabolism
- HSP70 Heat-Shock Proteins/genetics*
- HSP70 Heat-Shock Proteins/metabolism
- Membrane Proteins/genetics*
- Membrane Proteins/metabolism
- Phylogeny
- Up-Regulation
- PubMed
- 26988288 Full text @ Fish Shellfish Immunol.
Citation
Wang, X., Zhang, T., Mao, H., Mi, Y., Zhong, B., Wei, L., Liu, X., Hu, C. (2016) Grass carp (Ctenopharyngodon idella) ATF6 (Activating Transcription Factor 6) modulates the transcriptional level of GRP78 and GRP94 in CIK cells. Fish & shellfish immunology. 52:65-73.
Abstract
ATF transcription factors are stress proteins containing alkaline area-leucine zipper and play an important role in endoplasmic reticulum stress. ATF6 is a protective protein which regulates the adaptation of cells to ER stress by modulating the transcription of UPR (Unfolded Protein Response) target genes, including GRP78 and GRP94. In the present study, a grass carp (Ctenopharyngodon idella) ATF6 full-length cDNA (named CiATF6, KT279356) has been cloned and identified. CiATF6 is 4176 bp in length, comprising 159 nucleotides of 5'-untranslated sequence, a 1947 nucleotides open reading frame and 2170 nucleotides of 3'-untranslated sequences. The largest open reading frame of CiATF6 translates into 648 aa with a typical DNA binding domain (BRLZ domain) and shares significant homology to the known ATF6 counterparts. Phylogenetic reconstruction confirmed its closer evolutionary relationship with other fish counterparts, especially with Zebrafish ATF6. RT-PCR showed that CiATF6 was ubiquitously expressed and significantly up-regulated after stimulation with thermal stress in all tested grass carp tissues. In order to know more about the role of CiATF6 in ER stress, recombinant CiATF6N with His-tag was over-expressed in Rosetta Escherichia coli, and the expressed protein was purified by affinity chromatography with Ni-NTA His-Bind Resin. In vitro, gel mobility shift assays were employed to analyze the interaction of CiATF6 protein with the promoters of grass carp GRP78 and GRP94, respectively. The result has shown that CiATF6 could bind to these promoters with high affinity by means of its BRLZ mainly. To further study the transcriptional regulatory mechanism of CiATF6, Dual-luciferase reporter assays were applied. Recombinant plasmids of pGL3-GRP78P and pGL3-CiGRP94P were constructed and transiently co-transfected with pcDNA3.1-CiATF6 (pcDN3.1-CiATF6-nBRLZ, respectively) into C. idella kidney (CIK) cells. The result has shown that CiATF6 could activate CiGRP78 and CiGRP94 promoters.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping